US20040005664A1

US20040005664A1 - Novel 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 and 8843 molecules and uses therefor

Info

Publication number: US20040005664A1
Application number: US10/410,764
Authority: US
Inventors: Rachel Meyers; Kyle Macbeth; Rory Curtis; Laura Rudolph-Owen; Nadine Weich; Peter Olandt; Fong-Ying Tsai; Rosana Kapeller-Libermann; Joseph Carroll
Original assignee: Millennium Pharmaceuticals Inc
Current assignee: Millennium Pharmaceuticals Inc
Priority date: 2000-04-25
Filing date: 2003-04-10
Publication date: 2004-01-08
Also published as: US20100150901A1; US20070065848A1; US7485308B2

Abstract

The invention provides isolated nucleic acids molecules, designated 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 and 8843 nucleic acid molecules. The invention also provides antisense nucleic acid molecules, recombinant expression vectors containing 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 and 8843 nucleic acid molecules, host cells into which the expression vectors have been introduced, and nonhuman transgenic animals in which a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 gene has been introduced or disrupted. The invention still further provides isolated 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 proteins, fusion proteins, antigenic peptides and anti-26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 antibodies. Diagnostic and therapeutic methods utilizing compositions of the invention are also provided.

Description

RELATED APPLICATIONS

The present application is a continuation-in-part of U.S. patent application Ser. No. 09/924,358, filed Aug. 6, 2001 (pending), which claims the benefit of U.S. Provisional Application Serial No. 60/229,300, filed Sep. 1, 2000. The present application is also a continuation-in-part of U.S. patent application Ser. No. 10/350,553, filed Jan. 24, 2003 (pending), which claims the benefit of U.S. Provisional Application Serial No. 60/351,572, filed Jan. 24, 2002. The present application is also a continuation-in-part of U.S. patent application Ser. No. 09/966,614, filed Sep. 27, 2001 (pending), which claims the benefit of U.S. Provisional Application Serial No. 60/238,054, filed Oct. 5, 2000. The present application is also a continuation-in-part of U.S. patent application Ser. No. 10/281,094, filed Oct. 25, 2002 (pending), which claims the benefit of U.S. Provisional Application Serial No. 60/347,815, filed Oct. 29, 2001. The present application is also a continuation-in-part of U.S. patent application Ser. No. 10/076,535, filed Feb. 15, 2002 (pending), which claims the benefit of U.S. Provisional Application Serial No. 60/269,440, filed Feb. 16, 2001. The present application is also a continuation-in-part of U.S. patent application Ser. No. 09/860,352, filed May 17, 2001 (pending), which claims the benefit of U.S. Provisional Application Serial No. 60/205,301, filed May 19, 2000. The present application is also a continuation-in-part of U.S. patent application Ser. No. 09/593,927, filed Jun. 15, 2000 (pending), which claims the benefit of U.S. Provisional Application Serial No. 60/199,391, filed Apr. 25, 2000. The present application is also a continuation-in-part of U.S. patent application Ser. No. 10/226,410, filed Aug. 23, 2002 (pending), which claims the benefit of U.S. Provisional Application Serial No. 60/314,884, filed on Aug. 24, 2001. The present application is also a continuation-in-part of U.S. patent application Ser. No. 09/997,816, filed Nov. 29, 2001 (pending), which claims the benefit of U.S. Provisional Application Serial No. 60/250,186, filed Nov. 30, 2000. The present application is also a continuation-in-part of U.S. patent application Ser. No. 09/686,673, filed Oct. 11, 2000 (pending). The entire contents of each of the above-referenced patent applications are incorporated herein by this reference. [0001]

BACKGROUND OF THE INVENTION

The enormous variety of biochemical reactions that comprise life are nearly all mediated by a series of biological catalysts known as enzymes. Enzymes are proteins which possess specific catalytic activities that enable them to catalyze a series of reactions, hence enabling metabolic pathways to degrade and to reconstruct products needed to maintain organisms. By the binding of substrates through geometrically and physically complementary reactions, enzymes are stereospecific in binding substrates as well as in catalyzing reactions. The stringency for this stereospecificity varies as some enzymes are more specific to the identity of their substrates, while others are capable of binding multiple substrates and can catalyze numerous types of reactions.

Examples of enzymes include, for example, arginine methyltransferases, glycosyltransferases, gamma-glutamyltraspeptidases, phosphoribosylglycinamide transferases, acyltransferases, acyl-CoA dehydrogenases, fatty acid amide hydrolases, aminotransferases, zinc carboxypeptidases, protein kinases, DEAD helicases, short-chain dehydrogenase/reductases and phosphatases. Such enzymes have the ability, for example: 1) to transfer an activated sugar residue to an acceptor molecule; 2) to modulate the processing, folding, and secretion of proteins; 3) to transport amino acids in the form of their gamma-glutamyl derivatives; 4) to regulate the metabolism of glutathione; 5) to regulate the synthesis of purines; 6) to modulate cell division and proliferation; 7) to modulate cell death; 8) to transfer an acyl chain to a lipid precursor; 9) to regulate lipid biosynthesis; 10) to catalyze the transfer of hydrogen and electrons from one compound to another; 11) to catalyze the I,θ-dehydrogenation of fatty acyl-CoA derivatives; 12) to bind and catabolize fatty acid amides; 13) to modulate metabolism, e.g., amino acid metabolism; 14) to bind an amino acid, e.g., L-alanine; 15) to bind an oxo acid, e.g., pyruvate; 16) to modulate the formation of a zinc ion complex with a carbonyl group of a substrate polypeptide and polarization of the carbon-oxygen bond; 17) to modulate formation of a tetrahedral intermediate due to attack of the carbonyl carbon by water in a reaction assisted by a carboxylate side chain of glutamate; 18) to modulate the production of a dianion intermediate by rapid ionization of the tetrahedral intermediate produced; 19) to modulate ATP dependent nucleic acid unwinding; 20) to modulate RNA metabolism (e.g., nuclear transcription, and mRNA splicing); 21) to modulate steroid biosynthesis or metabolism (breakdown); 22) to catalyze the removal of a phosphate group attached to a tyrosine residue in a protein; 23) to catalyze the removal of a phosphate group attached to a serine or threonine residue in a protein; 24) to modulate an intracellular signaling pathway, e.g., a MAP kinase or ERK kinase pathway; 25) to regulate the transmission of signals from cellular receptors, e.g., cardiac cell growth factor receptors; as well as many others. Accordingly, there exists a need to identify additional human enzymes, for example, for use as disease markers and as targets for identifying various therapeutic modulators.

SUMMARY OF THE INVENTION

The present invention is based, at least in part, on the discovery of novel nucleic acid molecules and proteins encoded by such nucleic acid molecules, referred to herein as “26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843”. The 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 nucleic acid and protein molecules of the present invention are useful as modulating agents in regulating a variety of cellular processes, e.g., including cell proliferation, differentiation, growth and division. In particular, these nucleic acid molecules will be advantageous in the regulation of any cellular function, uncontrolled proliferation and differentiation, such as in cases of cancer. Accordingly, in one aspect, this invention provides isolated nucleic acid molecules encoding 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 proteins or biologically active portions thereof, as well as nucleic acid fragments suitable as primers or hybridization probes for the detection of 26199, 33530, 33949,47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843-encoding nucleic acids.

The nucleotide sequence of the cDNA encoding 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843, and the amino acid sequence of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 polypeptides are depicted in Table 1.

TABLE 1


Sequences of the invention

				ATCC
Gene				accession
Name	cDNA	Protein	Coding Region	number

26199	SEQ ID NO:1	SEQ ID NO:2	SEQ ID NO:3	N/A
33530	SEQ lD NO:4	SEQ ID NO:5	SEQ ID NO:6	PTA-3437
33949	SEQ ID NO:7	SEQ ID NO:8	SEQ ID NO:9	N/A
47148	SEQ ID NO:10	SEQ ID NO:11	SEQ ID NO:12	N/A
50226	SEQ ID NO:13	SEQ ID NO:14	SEQ ID NO:15	N/A
58764	SEQ ID NO:16	SEQ ID NO:17	SEQ ID NO:18	N/A
62113	SEQ ID NO:53	SEQ ID NO:54	SEQ ID NO:55	N/A
32144	SEQ ID NO:61	SEQ ID NO:62	SEQ ID NO:63	N/A
32235	SEQ ID NO:67	SEQ ID NO:68	SEQ ID NO:69	N/A
23565	SEQ ID NO:78	SEQ ID NO:79	SEQ ID NO:80	N/A
13305	SEQ ID NO:88	SEQ ID NO:89	SEQ ID NO:90	N/A
14911	SEQ ID NO:	SEQ ID NO:	SEQ ID NO:	PTA-3435
	100	101	102
86216	SEQ ID NO:	SEQ ID NO:	SEQ ID NO:	N/A
	113	114	115
25206	SEQ ID NO:	SEQ ID NO:	SEQ ID NO:	N/A
	122	123	124
8843	SEQ ID NO:	SEQ ID NO:	SEQ ID NO:	N/A
	129	130	131

Accordingly, in one aspect, the invention features a nucleic acid molecule, which encodes a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein or polypeptide, e.g., a biologically active portion of the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein. In a preferred embodiment, the isolated nucleic acid molecule encodes a polypeptide having the amino acid sequence of SEQ ID NO:2, 5, 8, 11, 14, 17, 54, 62, 68, 79, 89, 101, 114, 123 or 130. In other embodiments, the invention provides isolated 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 nucleic acid molecules having the nucleotide sequence shown in SEQ ID NO:1, 3, 4, 6, 7, 9, 10, 12, 13, 15, 16, 18, 53, 55, 61, 63, 67, 69, 78, 80, 88, 90, 100, 102, 113, 115, 122, 124, 129 or 131 or the nucleotide sequence of the DNA insert of the plasmid deposited with ATCC Accession Number PTA-3437 or PTA-3435. In still other embodiments, the invention provides nucleic acid molecules that are substantially identical (e.g., naturally occurring allelic variants) to the nucleotide sequence shown in SEQ ID NO:1, 3, 4, 6, 7, 9, 10, 12, 13, 15, 16, 18, 53, 55, 61, 63, 67, 69, 78, 80, 88, 90, 100, 102, 113, 115, 122, 124, 129 or 131 or the nucleotide sequence of the DNA insert of the plasmid deposited with ATCC Accession Number PTA-3437 or PTA-3435. In other embodiments, the invention provides a nucleic acid molecule which hybridizes under a stringent hybridization condition as described herein to a nucleic acid molecule comprising the nucleotide sequence of SEQ ID NO:1, 3, 4, 6, 7, 9, 10, 12, 13, 15, 16, 18, 53, 55, 61, 63, 67, 69, 78, 80, 88, 90, 100, 102, 113, 115, 122, 124, 129 or 131 or the nucleotide sequence of the DNA insert of the plasmid deposited with ATCC Accession Number PTA-3437 or PTA-3435, wherein the nucleic acid encodes a full length 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein or an active fragment thereof.

In a related aspect, the invention further provides nucleic acid constructs which include a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 nucleic acid molecule described herein. In certain embodiments, the nucleic acid molecules of the invention are operatively linked to native or heterologous regulatory sequences. Also included are vectors and host cells containing the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 nucleic acid molecules of the invention e.g., vectors and host cells suitable for producing polypeptides.

In another related aspect, the invention provides nucleic acid fragments suitable as primers or hybridization probes for the detection of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843-encoding nucleic acids.

In still another related aspect, isolated nucleic acid molecules that are antisense to a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 encoding nucleic acid molecule are provided.

In another aspect, the invention features 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 polypeptides, and biologically active or antigenic fragments thereof that are useful, e.g., as reagents or targets in assays applicable to treatment and diagnosis of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843-associated disorders. In another embodiment, the invention provides 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 polypeptides having a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 activity.

In other embodiments, the invention provides 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 polypeptides, e.g., a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 polypeptide having the amino acid sequence shown in SEQ ID NO:2, 5, 8, 11, 14, 17, 54, 62, 68, 79, 89, 101, 114, 123 or 130 or the amino acid sequence encoded by the cDNA insert of the plasmid deposited with ATCC Accession Number PTA-3437 or PTA-3435; an amino acid sequence that is substantially identical to the amino acid sequence shown in SEQ ID NO:2, 5, 8, 11, 14, 17, 54, 62, 68, 79, 89, 101, 114, 123 or 130 or the amino acid sequence encoded by the cDNA insert of the plasmid deposited with ATCC Accession Number PTA-3437 or PTA-3435; or an amino acid sequence encoded by a nucleic acid molecule having a nucleotide sequence which hybridizes under a stringent hybridization condition as described herein to a nucleic acid molecule comprising the nucleotide sequence of SEQ ID NO:1, 3, 4, 6, 7, 9, 10, 12, 13, 15, 16, 18, 53, 55, 61, 63, 67, 69, 78, 80, 88, 90, 100, 102, 113, 115, 122, 124, 129 or 131 or the nucleotide sequence of the insert of the plasmid deposited with ATCC Accession Number PTA-3437 or PTA-3435, wherein the nucleic acid encodes a full length 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein or an active fragment thereof.

In a related aspect, the invention further provides nucleic acid constructs which include a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 nucleic acid molecule described herein.

In a related aspect, the invention provides 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 polypeptides or fragments operatively linked to non-26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 polypeptides to form fusion proteins.

In another aspect, the invention features antibodies and antigen-binding fragments thereof, that react with, or more preferably specifically or selectively bind 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 polypeptides.

In another aspect, the invention provides methods of screening for compounds that modulate the expression or activity of the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 polypeptides or nucleic acids.

In still another aspect, the invention provides a process for modulating 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 polypeptide or nucleic acid expression or activity, e.g., using the compounds identified in the screens described herein. In certain embodiments, the methods involve treatment of conditions related to aberrant activity or expression of the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 polypeptides or nucleic acids, such as conditions or disorders involving aberrant or deficient 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 expression. Examples of such disorders include, but are not limited to cellular proliferative and/or differentiative disorders, brain disorders, platelet disorders, breast disorders, colon disorders, kidney (renal) disorders, lung disorders, ovarian disorders, prostate disorders, hematopoeitic disorders, pancreatic disorders, skeletal muscle disorders, skin (dermal) disorders, disorders associated with bone metabolism, immune, e.g., inflammatory, disorders, cardiovascular disorders, endothelial cell disorders, liver disorders, viral diseases, pain disorders, metabolic disorders, neurological or CNS disorders, erythroid disorders or anemic disorders.

The invention also provides assays for determining the activity of or the presence or absence of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 polypeptides or nucleic acid molecules in a biological sample, including for disease diagnosis.

In a further aspect, the invention provides assays for determining the presence or absence of a genetic alteration in a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 polypeptide or nucleic acid molecule, including for disease diagnosis.

In another aspect, the invention features a two dimensional array having a plurality of addresses, each address of the plurality being positionally distinguishable from each other address of the plurality, and each address of the plurality having a unique capture probe, e.g., a nucleic acid or peptide sequence. At least one address of the plurality has a capture probe that recognizes a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 molecule. In one embodiment, the capture probe is a nucleic acid, e.g., a probe complementary to a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 nucleic acid sequence. In another embodiment, the capture probe is a polypeptide, e.g., an antibody specific for 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 polypeptides. Also featured is a method of analyzing a sample by contacting the sample to the aforementioned array and detecting binding of the sample to the array.

Other features and advantages of the invention will be apparent from the following detailed description, and from the claims.

DETAILED DESCRIPTION OF THE INVENTION

Human 26199, 33530, 33949, 47148, 50226, and 58764

The present invention is based, in part, on the discovery of novel human transferase family members, referred to herein as “26199, 33530, 33949, 47148, 50226, and 58764”.

Human 26199

The human 26199 sequence (SEQ ID NO:1), which is approximately 1828 nucleotides long including untranslated regions, contains a predicted methionine-initiated coding sequence of about 687 nucleotides (nucleotides 56-742 of SEQ ID NO:1; 1-687 of SEQ ID NO:3, not including the terminal codon). The coding sequence encodes a 229 amino acid protein (SEQ ID NO:2). This mature protein form is approximately 229 amino acid residues in length (from about amino acid 1 to amino acid 229 of SEQ ID NO:2).

A BLAST alignment of human 26199 with a consensus amino acid sequence derived from a ProDomain “chromosome genomic DNA 5 FIS clone:MLN1 T6D22.22 UME3-HDA1 tumor-related ZHB0014.1” (PD113097; Release 2001.1) shows amino acid residues 2 to 115 of the 119 amino acid consensus sequence (SEQ ID NO:19) aligns with the “chromosome genomic DNA 5 FIS clone:MLN1 T6D22.22 UME3-HDA1 tumor-related ZHB0014.1” domain of human 26199, amino acid residues 7 to 120 of SEQ ID NO:2.

A BLAST alignment of human 26199 with a consensus amino acid sequence derived from a ProDomain “P1 genomic clone:MLN1 chromosome 5” (PD289255; Release 2001.1) shows amino acid residues 3 to 104 of the 111 amino acid consensus sequence (SEQ ID NO:20) aligns with the “P1 genomic clone:MLN1 chromosome 5” domain of human 26199, amino acid residues 123 to 226 of SEQ ID NO:2.

A BLAST alignment of human 26199 with a consensus amino acid sequence derived from a ProDomain “MRPL37-RIF1” (PD113089; Release 2001.1) shows amino acid residues 191 to 401 of the 419 amino acid consensus sequence (SEQ ID NO:21) aligns with the “MRPL37-RIF1” domain of human 26199, amino acid residues 15 to 208 of SEQ ID NO:2.

Human 26199 contains the following regions or other structural features: two predicted transmembrane domains (predicted by MEMSAT, Jones et al. (1994) Biochemistry 33:3038-3049) which extend from about amino acid residue 33-49 and 74-94 of SEQ ID NO:2; two glycosaminoglycan attachment sites (PS00002) located at about amino acids 59-62 and 76-79 of SEQ ID NO:2; one predicted cAMP- and cGMP-dependent protein kinase phosphorylation site (PS0004) located at about amino acids 222-225. of SEQ ID NO:2; two predicted protein kinase C phosphorylation sites (PS00005) located at about amino acids 67-69 and 158-160 of SEQ ID NO:2; six predicted casein kinase II phosphorylation sites (PS00006) located at about amino 7-10, 70-73, 95-98, 135-138, 158-161 and 163-166 of SEQ ID NO:2; four predicted N-myristoylation sites (PS00008) located at about amino acids 36-41, 75-80, 82-87 and 117-122 of SEQ ID NO:2; and one predicted prokaryotic membrane lipoprotein lipid attachment site (PS00013) located at about amino acids 30-40 of SEQ ID NO:2.

In one embodiment, a 26199 family member can include at least one and preferably two transmembrane domains. Furthermore, a 26199 family member can include at least one and preferably two glycosaminoglycan attachment sites (PS00002); at least one cAMP- and cGMP-dependent protein kinase phosphorylation site (PS00004); at least one, and preferably two protein kinase C phosphorylation sites (PS00005); at least one, two, three, four, five, and preferably six casein kinase II phosphorylation sites (PS00006); at least one, two, three, and preferably four N-myristolyation sites (PS00008); at least one prokaryotic membrane lipoprotein lipid attachment site (PS00013).

26199 is overexpressed in human breast and lung carcinomas. It is expected that inhibition of this arginine methyltransferase will inhibit tumor progression.

For general information regarding PFAM identifiers, PS prefix and PF prefix domain identification numbers, refer to Sonnhammer et al. (1997) Protein 28:405-420.

A hydropathy plot of human 26199 was performed. Polypeptides of the invention include fragments which include: all or part of a hydrophobic sequence, e.g., the sequence from about amino acid 40 to 50, from about 80 to 100, and from about 135 to 145 of SEQ ID NO:2; all or part of a hydrophilic sequence, e.g., the sequence from about amino acid 50 to 70, from about 170 to 190, and from about 200 to 210 of SEQ ID NO:2; a sequence which includes a Cys, or a glycosylation site.

Human 33530

The human 33530 sequence (SEQ ID NO:4), which is approximately 1408 nucleotides long including untranslated regions, contains a predicted methionine-initiated coding sequence of about 1248 nucleotides (nucleotides 36-1283 of SEQ ID NO:4; 1-1248 of SEQ ID NO:6, not including the terminal codon). The coding sequence encodes a 416 amino acid protein (SEQ ID NO:5). This mature protein form is approximately 416 amino acid residues in length (from about amino acid 1 to amino acid 416 of SEQ ID NO:5).

An alignment of the “glycosyl transferase group 1” domain of human 33530 with a consensus amino acid sequence derived from a hidden Markov model (HMM) from PFAM shows the consensus amino acid sequence (SEQ ID NO:22) aligns with amino acids 211 to 393 of SEQ ID NO:5.

A BLAST alignment of human 33530 with a consensus amino acid sequence derived from a ProDomain “a similar BA13B9.1 glycosyltransferase novel cDNA MNCB-5081 brain” (PD346441; Release 2001.1) shows amino acid residues 1 to 49 of the 49 amino acid consensus sequence (SEQ ID NO:23) aligns with the “a similar BA13B9.1 glycosyltransferase novel cDNA MNCB-5081 brain” domain of human 33530, amino acid residues 367 to 415 of SEQ ID NO:5.

A BLAST alignment of human 33530 with a consensus amino acid sequence derived from a ProDomain “glycosyltransferase ALG2 similar musculus F9K20.16 other novel brain 2.4.1.” (PD011566; Release 2001.1) shows amino acid residues 4 to 84 of the 84 amino acid consensus sequence (SEQ ID NO:24) aligns with the “glycosyltransferase ALG2 similar musculus F9K20.16 other novel brain 2.4.1.” domain of human 33530, amino acid residues 17 to 95 of SEQ ID NO:5.

A BLAST alignment of human 33530 with a consensus amino acid sequence derived from a ProDomain “transferase glycosyltransferase biosynthesis lipopolysaccharide galactosyltransferase glucosyltransferase mannosyl 2.4.1.-mannosyltransferase” (PD010528; Release 2001.1) shows amino acid residues 15 to 158 of the 164 amino acid consensus sequence (SEQ ID NO:25) aligns with the “transferase glycosyltransferase biosynthesis lipopolysaccharide galactosyltransferase glucosyltransferase mannosyl 2.4.1.-mannosyltransferase” domain of human 33530, amino acid residues 280 to 413 of SEQ ID NO:5.

A BLAST alignment of human 33530 with a consensus amino acid sequence derived from a ProDomain “F9K20.16” (PD241981; Release 2001.1) shows amino acid residues 1 to 46 of the 46 amino acid consensus sequence (SEQ ID NO:26) aligns with the “F9K20.16” domain of human 33530, amino acid residues 96 to 143 of SEQ ID NO:5.

A BLAST alignment of human 33530 with a consensus amino acid sequence derived from a ProDomain “glycosyltransferase 2.4.1.-ALG2 transmembrane glycoprotein” (PD258606; Release 2001.1) shows amino acid residues 15 to 60 of the 60 amino acid consensus sequence (SEQ ID NO:27) aligns with the “glycosyltransferase 2.4.1.-ALG2 transmembrane glycoprotein” domain of human 33530, amino acid residues 109 to 155 of SEQ ID NO:5.

A BLAST alignment of human 33530 with a consensus amino acid sequence derived from a ProDomain “glycosyltransferase” (PD309959; Release 2001.1) shows amino acid residues 5 to 161 of the 199 amino acid consensus sequence (SEQ ID NO:28) aligns with the “glycosyltransferase” domain of human 33530, amino acid residues 216 to 382 of SEQ ID NO:5.

Human 33530 contains the following regions or other structural features: one predicted glycosyl transferase group 1 domain (PFAM Accession Number PF00534) located at about amino acid residues 211-393 of SEQ ID NO:5; one predicted transmembrane domain (predicted by MEMSAT, Jones et al. (1994) Biochemistry 33:3038-3049) which extends from about amino acid residue 85-105 of SEQ ID NO:5; two predicted N-glycosylation sites (PS00001) located at about amino acids 204-207 and 239-242 of SEQ ID NO:5; one predicted cAMP- and cGMP-dependent protein kinase phosphorylation site (PS0004) located at about amino acids 146-149 of SEQ ID NO:5; five predicted protein kinase C phosphorylation sites (PS00005) located at about amino acids 46-48, 145-147, 187-189, 304-306 and 381-383 of SEQ ID NO:5; five predicted casein kinase II phosphorylation sites (PS00006) located at about amino 145-148, 192-195, 206-209, 255-258 and 302-305 of SEQ ID NO:5; five predicted N-myristoylation sites (PS00008) located at about amino acids 25-30, 78-83, 85-90, 168-173 and 294-299 of SEQ ID NO:5; and one predicted amidation site (PS00009) located at about amino acids 222-225 of SEQ ID NO:5.

In one embodiment, a 33530 family member can include at least one glycosyl transferase group 1 domain (PFAM Accession Number PF00534) and at least one transmembrane domain. Furthermore, a 33530 family member can include at least one and preferably two N-glycosylation sites (PS00001); at least one cAMP- and cGMP-dependent protein kinase phosphorylation site (PS00004); at least one, two, three, four, and preferably five protein kinase C phosphorylation sites (PS00005); at least one, two, three, four, and preferably five casein kinase II phosphorylation sites (PS00006); at least one, two, three, four, and preferably five N-myristolyation sites (PS00008); at least one amidation site (PS00009).

A hydropathy plot of human 33530 was performed. Polypeptides of the invention include fragments which include: all or part of a hydrophobic sequence, e.g., the sequence from about amino acid 30 to 45, from about 85 to 105, and from about 115 to 125 of SEQ ID NO:5; all or part of a hydrophilic sequence, e.g., the sequence from about amino acid 55 to 70, from about 155 to 160, and from about 270 to 290 of SEQ ID NO:5; a sequence which includes a Cys, or a glycosylation site.

A plasmid containing the nucleotide sequence encoding human 33530 was deposited with American Type Culture Collection (ATCC), 10801 University Boulevard, Manassas, Va. 20110-2209, on Jun. 7, 2001 and assigned Accession Number PTA-3437. This deposit will be maintained under the terms of the Budapest Treaty on the International Recognition of the Deposit of Microorganisms for the Purposes of Patent Procedure. This deposit was made merely as a convenience for those of skill in the art and is not an admission that a deposit is required under 35 U.S.C. §112.

Human 33949

The human 33949 sequence (SEQ ID NO:7), which is approximately 2327 nucleotides long including untranslated regions, contains a predicted methionine-initiated coding sequence of about 1824 nucleotides (nucleotides 148-1971 of SEQ ID NO:7; 1-1824 of SEQ ID NO:9, not including the terminal codon). The coding sequence encodes a 608 amino acid protein (SEQ ID NO:8). The protein contains a signal sequence from about amino acid 1 to 37 of SEQ ID NO:8. The mature protein form is approximately 571 amino acid residues in length (from about amino acid 38 to amino acid 608 of SEQ ID NO:8).

An alignment of the “glycosyl transferase group 2” domain of human 33949 with a consensus amino acid sequence derived from a hidden Markov model (HMM) from PFAM shows the consensus amino acid sequence (SEQ ID NO:29) aligns with amino acids 154 to 341 of SEQ ID NO:8.

An alignment of the “QXW lectin repeat (Ricin_B_lectin)” domain of human 33949 with a consensus amino acid sequence derived from a hidden Markov model (HMM) from PFAM shows the consensus amino acid sequence (SEQ ID NOS: 30-32) aligns with amino acids 483 to 526, 527 to 567 and 568 to 606 of SEQ ID NO:8.

A BLAST alignment of human 33949 with a consensus amino acid sequence derived from a ProDomain “WUGSC:H_DJ0981007.2 cDNA: FIS COL08230 FLJ21634” (PD354231; Release 2001.1) shows amino acid residues 1 to 102 of the 102 amino acid consensus sequence (SEQ ID NO:33) aligns with the “WUGSC:H_DJ0981007.2 cDNA: FIS COL08230 FLJ21634” domain of human 33949, amino acid residues 1 to 102 of SEQ ID NO:8.

A BLAST alignment of human 33949 with a consensus amino acid sequence derived from a ProDomain “acetylgalactosaminyltransferase N-acetylgalactosaminyltransferase polypeptide UDP-GALNAC:polypeptide protein-glyco glycosyltransferase” (PD003677; Release 2001.1) shows amino acid residues 2 to 130 of the 130 amino acid consensus sequence (SEQ ID NO:34) aligns with the “acetylgalactosaminyltransferase N-acetylgalactosaminyltransferase polypeptide UDP-GALNAC:polypeptide protein-glyco glycosyltransferase” domain of human 33949, amino acid residues 103 to 229 of SEQ ID NO:8.

A BLAST alignment of human 33949 with a consensus amino acid sequence derived from a ProDomain “acetylgalactosaminyltransferase N-acetylgalactosaminyltransferase polypeptide UDP-GALNAC:polypeptide protein-FIS GALNAC-T1” (PD003162; Release 2001.1) shows amino acid residues 1 to 62 of the 62 amino acid consensus sequence (SEQ ID NO:35) aligns with the “acetylgalactosaminyltransferase N-acetylgalactosaminyltransferase polypeptide UDP-GALNAC:polypeptide protein-FIS GALNAC-T1” domain of human 33949, amino acid residues 347 to 406 of SEQ ID NO:8.

A BLAST alignment of human 33949 with a consensus amino acid sequence derived from a ProDomain “FIS cDNA: WUGSC:H_DJ0981007.2 HRC08167 COL08230 FLJ21634 FLJ22403” (PD334332; Release 2001.1) shows amino acid residues 1 to 41 and 2 to 37 of the 41 amino acid consensus sequence (SEQ ID NOs: 36-37) aligns with the “FIS cDNA: WUGSC:H_DJ0981007.2 HRC08167 COL08230 FLJ21634 FLJ22403” domain of human 33949, amino acid residues 568 to 608 and 484 to 521 of SEQ ID NO:8.

A BLAST alignment of human 33949 with a consensus amino acid sequence derived from a ProDomain “N-acetylgalactosaminyltransferase polypeptide UDP-GALNAC:polypeptide protein-glyco glycosyltransferase” (PD301297; Release 2001.1) shows amino acid residues 1 to 80 of the 80 amino acid consensus sequence (SEQ ID NO:38) aligns with the “N-acetylgalactosaminyltransferase polypeptide UDP-GALNAC:polypeptide protein-glyco glycosyltransferase” domain of human 33949, amino acid residues 273 to 346 of SEQ ID NO:8.

Human 33949 contains the following regions or other structural features: one predicted glycosyl transferase group 2 domain located at about amino acid residues 154-341 of SEQ ID NO:8; three predicted transmembrane domains (predicted by MEMSAT, Jones et al. (1994) Biochemistry 33:3038-3049) which extends from about amino acid residues 8-28, 150-168 and 268-284 of SEQ ID NO:8; two predicted N-glycosylation site (PS00001) located at about amino acids 29-32 and 428-431 of SEQ ID NO:8; eleven predicted protein kinase C phosphorylation sites (PS00005) located at about amino acids 5-7, 51-53, 124-126, 220-222, 358-360, 399-401, 416-418, 430-432, 443-445, 490-492 and 501-503 of SEQ ID NO:8; six predicted casein kinase II phosphorylation sites (PS00006) located at about amino 82-85, 173-176, 193-196, 220-223, 246-249 and 345-348 of SEQ ID NO:8; one predicted tyrosine kinase phosphorylation site (PS00007) located at about amino acids 445-452 of SEQ ID NO:8; and nine predicted N-myristoylation sites (PS00008) located at about amino acids 12-17, 99-104, 224-229, 232-237, 327-332, 341-346, 387-392, 555-560 and 586-591 of SEQ ID NO:8.

In one embodiment, a 33949 family member can include at least one glycosyl transferase group 2 domain (PFAM Accession Number PF00535) and at least one, two and preferably three transmembrane domains. Furthermore, a 33949 family member can include at least one and preferably two N-glycosylation sites (PS00001); at least one, two, three, four, five, six, seven, eight, nine, ten and preferably eleven protein kinase C phosphorylation sites (PS00005); at least one, two, three, four, five and preferably six casein kinase II phosphorylation sites (PS00006); at least one predicted tyrosine kinase phosphorylation site (PS00007); at least one, two, three, four, five, six, seven, eight, and preferably nine N-myristolyation sites (PS00008).

A hydropathy plot of human 33949 was performed. Polypeptides of the invention include fragments which include: all or part of a hydrophobic sequence, e.g., the sequence from about amino acid 235 to 245, from about 275 to 285, and from about 360 to 375 of SEQ ID NO:8; all or part of a hydrophilic sequence, e.g., the sequence from about amino acid 50 to 70, from about 130 to 150, and from about 385 to 400 of SEQ ID NO:8; a sequence which includes a Cys, or a glycosylation site.

The 33530 and 33949 proteins contain a significant number of structural characteristics in common with members of the glycosyltransferase family. A 33530 or 33949 polypeptide can include a “glycosyltransferase domain” or regions homologous with a “glycosyltransferase domain.”

As used herein, the term “glycosyltransferase” includes a protein or polypeptide which is capable of catalyzing the synthesis of glycoconjugates, including glycolipids, glycoproteins, and polysaccharides, by transferring an activated mono- or oligosaccharide residue to an existing acceptor molecule for the initiation or elongation of the carbohydrate chain. The acceptor can be a lipid, a protein, a heterocyclic compound, or another carbohydrate residue. Glycosyltransferases can be divided into numerous subfamilies based upon their specificity for sugar moieties and acceptor molecules. The glycosyltransferase domain of human 33530 bears similarity to a subfamily designated “group 1” glycosyltransferases. Members of this family transfer activated sugars to a variety of substrates, including glycogen, fructose-6-phosphate and lipopolysaccharides. Members of this family transfer UDP, ADP, GDP or CMP linked sugars. The glycosyltransferase domain of human 33949 bears similarity to a subfamily designated “group 2” glycosyltransferases. These enzymes comprise a diverse subfamily, whose members transfer sugar from UDP-glucose, UDP-N-acetyl-galactosamine, GDP-mannose or CDP-abequose, to a range of substrates including cellulose, dolichol phosphate and teichoic acids. Based on the sequence similarities, the 33530 or 33949 molecules of the present invention are predicted to have similar biological activities as glycosyltransferase family members.

Glycosyltransferases play roles in diverse cellular processes. For example, the major target of the natural IgM and IgG antibodies during hyperacute xenograft rejection is the terminal carbohydrate epitope Gal alpha(1,3)Gal, formed by the alpha 1,3galactosyl transferase, which places a terminal galactose residue in an alpha-linkage to another galactose (Sandrin et al. (1994) Immunol Rev 141:169-90). As another example, mutations in the Piga gene, the protein product of which mediates N-acetylglucosamine attachment to phosphatidylinositol, results in the clonal hematologic disorder, paroxysmal nocturnal hemoglobinuria (Ware et al. (1994) Blood 83:2418-22). Additionally, UDP-galactose:ceramide galactosyltransferase is the enzyme responsible for the biosynthesis of galactosylceramide, a molecule thought to play a critical role in myelin formation, signal transduction, viral and microbial adhesion, and oligodendrocyte development (Kapitonov et al. (1999) Glycobiology 9:961-78).

Glycosylation of glycoproteins and glycolipids is one of many molecular changes that accompany malignant transformation. GlcNAc-branched N-glycans and terminal Lewis antigen sequences have been observed to increase in some cancers, and to correlate with poor prognosis (Dennis et al. (1999) Biochim Biophys Acta 1473:21-34). Cellular membrane over-expression and shedding of acidic glycosphingolipids into the interstitial spaces and blood of cancer patients may play a central role in increased tumour cell growth, lack of immune cell recognition and neovascularization and could represent a molecular target for cancer therapy (Fish (1996) Med Hypotheses 46:140-44). Thus, the 33530 or 33949 molecules of the present invention may be involved in: 1) the transfer of an activated sugar residue to an acceptor molecule; 2) the processing, folding, and secretion of proteins; 3) the modulation of tumor cell growth and invasion; 4) myelin formation; 5) signal transduction; 6) viral and microbial adhesion; 7) oligodendrocyte development; 8) sperm-egg binding; 9) evasion of immune detection; 10) xenograft rejection; and 11) the ability to antagonize or inhibit, competitively or non-competitively, any of 1-11. Thus, the 33530 and 33949 molecules can act as novel diagnostic targets and therapeutic agents for controlling glycosyltransferase-related disorders, for example, such as those diseases associated with the activities described above. As the 33530 and 33949 molecules have homology to known glycosyltransferases, they are expected to be involved in controlling similar disorders.

33530 has been shown to be overexpressed in some human breast, lung and colon carcinomas, and underexpressed in some ovary and brain carcinomas. As such, inhibition of this gycosyltransferase may inhibit tumor progression in breast, lung and colon. Further, activation of this gycosyltransferase may inhibit tumor progression in ovary and brain.

The 33949 molecules also have similarities to bovine and murine N-acetygalactosaminyltransferase. Thus, without being bound by theory, the 33949 transferase, may be a human analogue of the bovine or murine N-acetygalactosaminyltransferase.

Further, 33949 is overexpressed in a subset of breast, ovary, lung and colon tumors. As such, inhibition of this N-acetylgalactosaminyltransferase may inhibit tumor progression.

33949 is clearly a member of the GalNAc-transferase family of glycosyl transferase type 2 enzymes. The overall sequence identity is quite high, and all of the residues known to be required for catalytic activity are present in 33949. In the lectin domain of the protein, which has been shown to be involved in glycopeptide substrate specifity, 33949 has a V where the majority of known active enzymes have a D (in the CLD motif). In one study with GalNAc-T1, this D was changed to an H and the enzyme was still active (albeit with 42% of maximum activity).

Phylogenetic analysis of 33949 indicates that both the catalytic and lectin domains may be most similar to the GalNAc-T6 and -T7 enzymes.

It is expected that 33949 will encode an active enzyme. Identification of the ‘natural’ protein substrate may not necessary for assay configuration since many GalNAc-transferases have been shown to work on various peptide substrates derived from mucin and other proteins.

As used herein, the term “glycosyltransferase domain” includes an amino acid sequence of about 100-250 amino acid residues in length and having a bit score for the alignment of the sequence to the glycosyltransferase domain (HMM) of at least 30. Preferably, a glycosyltransferase domain includes at least about 120-220 amino acids, more preferably about 120-200 amino acid residues, or about 130-180 amino acids and has a bit score for the alignment of the sequence to the glycosyltransferase domain (HMM) of at least 50 or greater. Glycosyltransferase domains (HMM) have been assigned numerous PFAM Accession Numbers, including PF00534 (group 1) and PF00535 (group 2). The glycosyltransferase domain (amino acids 211 to 393 of SEQ ID NO:5) of human 33530 aligns with a consensus amino acid sequence (group 1 glycosyltransferases) derived from a hidden Markov model. The glycosyltransferase domain (amino acids 154 to 341 of SEQ ID NO:8) of human 33949 aligns with a consensus amino acid sequence (group 2 glycosyltransferases) derived from a hidden Markov model.

In a preferred embodiment a 33530 or 33949 polypeptide or protein has a “glycosyltransferase domain” or a region which includes at least about 120-220 more preferably about 120-200 or 130-180 amino acid residues and has at least about 70% 80% 90% 95%, 99%, or 100% homology with a “glycosyltransferase domain,” e.g., the glycosyltransferase domain of human 33530 or 33949 (e.g., residues 211 to 393 of SEQ ID NO:5 or residues 154 to 341 of SEQ ID NO:8).

To identify the presence of a “glycosyltransferase” domain in a 33530 or 33949 protein sequence, and make the determination that a polypeptide or protein of interest has a particular profile, the amino acid sequence of the protein can be searched against a database of HMMs

Human 47148

The human 47148 sequence (SEQ ID NO:10), which is approximately 2172 nucleotides long including untranslated regions, contains a predicted methionine-initiated coding sequence of about 1986 nucleotides (nucleotides 31-2016 of SEQ ID NO:10; 1-1986 of SEQ ID NO:12, not including the terminal codon). The coding sequence encodes a 662 amino acid protein (SEQ ID NO:11). This mature protein form is approximately 662 amino acid residues in length (from about amino acid 1 to amino acid 662 of SEQ ID NO:11).

An alignment of the gamma-glutamyltranspeptidase domain of human 47148 with a consensus amino acid sequence derived from a hidden Markov model (HMM) from PFAM shows the consensus amino acid sequence (SEQ ID NO:39) aligns with amino acids 154 to 656 of SEQ ID NO:11.

A BLAST alignment of human 47148 with a consensus amino acid sequence derived from a ProDomain “FGENESH repeat novel gamma-glutamyltranspeptidase locus CCA D20S101 similar predictions. DJ18C9.2” (PD297327; Release 2001.1) shows amino acid residues 1 to 135 of the 135 amino acid consensus sequence (SEQ ID NO:40) aligns with the “FGENESH repeat novel gamma-glutamyltranspeptidase locus CCA D20S101 similar predictions. DJ18C9.2” domain of human 47148, amino acid residues 1 to 135 of SEQ ID NO:11.

A BLAST alignment of human 47148 with a consensus amino acid sequence derived from a ProDomain “gamma-glutamyltranspeptidase transferase acyltransferase precursor zymogen glutathione biosynthesis acylase glycoprotein” (PD127336; Release 2001.1) shows amino acid residues 2 to 294 of the 304 amino acid consensus sequence (SEQ ID NO:41) aligns with “gamma-glutamyltranspeptidase transferase acyltransferase precursor zymogen glutathione biosynthesis acylase glycoprotein” domain of human 47148, amino acid residues 200 to 471 of SEQ ID NO:11.

A BLAST alignment of human 47148 with a consensus amino acid sequence derived from a ProDomain “FGENESH repeat novel gamma-glutamyltranspeptidase locus CCA D20S101 similar predictions. DJ18C9.2” (PD290211; Release 2001.1) shows amino acid residues 1 to 114 of the 114 amino acid consensus sequence (SEQ ID NO:42) aligns with the “FGENESH repeat novel gamma-glutamyltranspeptidase locus CCA D20S101 similar predictions. DJ18C9.2” domain of human 47148, amino acid residues 549 to 662 of SEQ ID NO:11.

Human 47148 contains the following regions or other structural features: one predicted gamma-glutamyltranspeptidase domain (PFAM Accession Number PF01019) located at about amino acid residues 154-656 of SEQ ID NO:11; two predicted transmembrane domains (predicted by MEMSAT, Jones et al. (1994) Biochemistry 33:3038-3049) which extend from about amino acid residues 106-127 and 168-192 of SEQ ID NO:11; ten predicted N-glycosylation sites (PS00001) located at about amino acids 198-201, 267-270, 283-286, 330-333, 353-356, 394-397, 452-455, 519-522, 523-526 and 586-589 of SEQ ID NO:11; one predicted glycosaminoglycan attachment site (PS00002) located at about amino acids 182-185 of SEQ ID NO:11; seven predicted protein kinase C phosphorylation sites (PS00005) located at about amino acids 64-66, 88-90, 101-103, 285-287, 295-297, 411-413 and 638-640 of SEQ ID NO:11; ten predicted casein kinase II phosphorylation sites (PS00006) located at about amino 17-20, 56-59, 73-76, 88-91, 162-165, 347-350, 430-433, 434-437, 440-443 and 612-615 of SEQ ID NO:11; one predicted tyrosine kinase phosphorylation site (PS00007) located at about amino acids 421-427 of SEQ ID NO:11; fourteen predicted N-myristoylation sites (PS00008) located at about amino acids 78-83, 120-125, 140-145, 183-188, 227-232, 234-239, 328-333, 343-348, 364-369, 469-474, 505-510, 553-558, 562-567 and 637-642 of SEQ ID NO:11; and two predicted amidation sites (PS00009) located at about amino acids 42-45 and 535-538 of SEQ ID NO:11.

In one embodiment, a 47148 family member can include at least one gamma-glutamyltranspeptidase domain (PFAM Accession Number PF01019) and at least one and preferably two transmembrane domain. Furthermore, a 47148 family member can include at least one, two, three, four, five, six, seven, eight, nine, and preferably ten N-glycosylation sites (PS00001); at least one predicted glycosaminoglycan attachment site (PS00002); at least one, two, three, four, five, six, and preferably seven protein kinase C phosphorylation sites (PS00005); at least one, two, three, four, five, six, seven, eight, nine, and preferably ten casein kinase II phosphorylation sites (PS00006); at least one predicted tyrosine kinase phosphorylation site (PS00007); at least one, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen and preferably fourteen N-myristolyation sites (PS00008); at least one and preferably two amidation sites (PS00009).

A hydropathy plot of human 47148 was performed. Polypeptides of the invention include fragments which include: all or part of a hydrophobic sequence, e.g., the sequence from about amino acid 110 to 130, from about 165 to 180, and from about 480 to 490 SEQ ID NO:11; all or part of a hydrophilic sequence, e.g., the sequence from about amino acid 70 to 90, from about 210 to 225, and from about 520 to 540 of SEQ ID NO:11; a sequence which includes a Cys, or a glycosylation site.

Gamma-glutamyltraspeptidase plays an important role in the metabolism of glutathione. Located at the external surface of epithelial cells, gamma-glutamyltraspeptidase initiates extracellular glutathione breakdown, provides cells with local cysteine supply and contributes to maintain intracellular glutathione level. Gamma-glutamyltraspeptidase expression, highly sensitive to oxidative stress, is a part of the cell antioxidant defense mechanisms. Chikhi, N., et al. (1999) Comp Biochem Physiol B Biochem Mol Biol 122(4):367-80. Glutathione plays an essential role in protecting the pulmonary system for toxic insults (Potdar, P. D., et al. (1997) Am J Physiol 273(5 Pt 1):L1082-9). Thus, the 47148 molecules of the present invention may be involved in: 1) transport of amino acids in the form of their gamma-glutamyl derivatives; 2) metabolism of glutathione; 3) maintenance of cellular cysteine levels; 4) maintenance of intracellular glutathione levels; 5) metabolism of amino acids; and 6) the ability to antagonize or inhibit, competitively or non-competitively, any of 1-5. Thus, the 47148 molecules can act as novel diagnostic targets and therapeutic agents for controlling gamma-glutamyltraspeptidase-related disorders, for example, such as those diseases (e.g. liver disease) associated with the activities described above. As the 47148 molecules have homology to known gamma-glutamyltraspeptidase, they are expected to be involved in controlling similar disorders.

Gamma-glutamyltraspeptidase is conserved among species (Chikhi, supra) and, thus without being bound by theory, the 47148 gamma-glutamyltraspeptidase may be a human analogue of rat, mouse, or pig gamma-glutamyltraspeptidase.

As used herein, the term “gamma-glutamyltraspeptidase domain” includes an amino acid sequence of about 100-500 amino acid residues in length and having a bit score for the alignment of the sequence to the gamma-glutamyltraspeptidase domain (HMM) of at least 30. Preferably, a gamma-glutamyltraspeptidase domain includes at least about 200-500 amino acids, more preferably about 300-500 amino acid residues, or about 400-500 amino acids and has a bit score for the alignment of the sequence to the gamma-glutamyltraspeptidase domain (HMM) of at least 50 or greater. The gamma-glutamyltraspeptidase domain (HMM) has been assigned PFAM Accession Numbers, including PF01019. The gamma-glutamyltranspeptidase domain (amino acids 154 to 656 of SEQ ID NO:11) of human 47148 aligns with a consensus amino acid sequence derived from a hidden Markov model.

In a preferred embodiment a 47148 polypeptide or protein has a gamma-glutamyltraspeptidase domain” or a region which includes at least about 200-500 more preferably about 300-500 or 400-500 amino acid residues and has at least about 70% 80% 90% 95%, 99%, or 100% homology with a “gamma-glutamyltraspeptidase domain,” e.g., the gamma-glutamyltraspeptidase domain of human 47148 (e.g., residues 154 to 656 of SEQ ID NO:11).

To identify the presence of a “gamma-glutamyltraspeptidase” domain in a 47148 protein sequence, and make the determination that a polypeptide or protein of interest has a particular profile, the amino acid sequence of the protein can be searched against a database of HMMs

Human 50226

The human 50226 sequence (SEQ ID NO:13), which is approximately 1252 nucleotides long including untranslated regions, contains a predicted methionine-initiated coding sequence of about 1167 nucleotides (nucleotides 18-1184 of SEQ ID NO:13; 1-1167 of SEQ ID NO:15, not including the terminal codon). The coding sequence encodes a 389 amino acid protein (SEQ ID NO:14). The protein contains a signal sequence from about amino acid 1 to 17 of SEQ ID NO:14. The mature protein form is approximately 372 amino acid residues in length (from about amino acid 18 to amino acid 389 of SEQ ID NO:14).

An alignment of the formyl transferase domain of human 50226 with a consensus amino acid sequence derived from a hidden Markov model (HMM) from PFAM shows the consensus amino acid sequence (SEQ ID NO:43) aligns with amino acids 119 to 220 of SEQ ID NO:14.

A BLAST alignment of human 50226 with a consensus amino acid sequence derived from a ProDomain “transferase formyltransferase phosphoribosylglycinamide biosynthesis methionyl-tRNA methyltransferase purine transformylase formyltetrahydrofolate hydrolase” (PD001209; Release 2001.1) shows amino acid residues 42 to 149 of the 156 amino acid consensus sequence (SEQ ID NO:44) aligns with the “transferase formyltransferase phosphoribosylglycinamide biosynthesis methionyl-tRNA methyltransferase purine transformylase formyltetrahydrofolate hydrolase” domain of human 50226, amino acid residues 117 to 221 of SEQ ID NO:14.

A BLAST alignment of human 50226 with a consensus amino acid sequence derived from a ProDomain “formyltransferase methionyl-tRNA methyltransferase biosynthesis one-carbon metabolism 10-formyltetrahydrofolate 10-FTHFDH dehydrogenase” (PD004966; Release 2001.1) shows amino acid residues 10 to 123 of the 129 amino acid consensus sequence (SEQ ID NO:45) aligns with the “formyltransferase methionyl-tRNA methyltransferase biosynthesis one-carbon metabolism 10-formyltetrahydrofolate 10-FTHFDH dehydrogenase” domain of human 50226, amino acid residues 238 to 355 of SEQ ID NO:14.

Human 50226 contains the following regions or other structural features: one predicted formyl transferase domain (PFAM Accession Number PF00551) located at about amino acid residues 119-220 of SEQ ID NO:14; one predicted N-glycosylation site (PS00001) located at about amino acids 292-295 of SEQ ID NO:14; five predicted protein kinase C phosphorylation sites (PS00005) located at about amino acids 90-92, 200-202, 282-284, 369-371 and 374-376 of SEQ ID NO:14; two predicted casein kinase II phosphorylation sites (PS00006) located at about amino 200-203 and 341-344 of SEQ ID NO:14; two predicted N-myristoylation sites (PS00008) located at about amino acids 16-21 and 121-126 of SEQ ID NO:14; and one predicted leucine zipper pattern (PS00029) located at about amino acids 129-150 of SEQ ID NO:14.

In one embodiment, a 50226 family member can include at least one formyl transferase domain (PFAM Accession Number PF00551). Furthermore, a 50226 family member can include at least one N-glycosylation site (PS00001); at least one, two, three, four, and preferably five protein kinase C phosphorylation sites (PS00005); at least one, and preferably two casein kinase II phosphorylation sites (PS00006); at least one, and preferably two N-myristolyation sites (PS00008); at least one leucine zipper pattern (PS00029).

A hydropathy plot of human 50226 was performed. Polypeptides of the invention include fragments which include: all or part of a hydrophobic sequence, e.g., the sequence from about amino acid 122 to 130, from about 140 to 150, and from about 285 to 300 SEQ ID NO:14; all or part of a hydrophilic sequence, e.g., the sequence from about amino acid 35 to 50, from about 230 to 240, and from about 350 to 370 of SEQ ID NO:14; a sequence which includes a Cys, or a glycosylation site.

The 50226 protein has similarities to formyl transferase, specifically, phosphoribosylglycinamide transferase, which plays a role in the de novo purine biosynthetic pathway. Thus, the 50226 molecules of the present invention may be involved in: 1) synthesis of purines; 2) modulation of cell division and proliferation; 3) the modulation of cell death; and 4) the ability to antagonize or inhibit, competitively or non-competitively, any of 1-3. Thus, the 50226 molecules can act as novel diagnostic targets and therapeutic agents for controlling phosphoribosylglycinamide transferase-related disorders, for example, such as those diseases (e.g. cancer) associated with the activities described above. As the 50226 molecules have homology to known phosphoribosylglycinamide transferase, they are expected to be involved in controlling similar disorders.

Phosphoribosylglycinamide transferase is conserved among species and, thus without being bound by theory, the 50226 phosphoribosylglycinamide transferase may be a human analogue of chicken or mouse phosphoribosylglycinamide transferase.

50226 has been shown to be overexpressed in some human breast, lung and colon carcinomas, and underexpressed in some ovary carcinomas. As such, inhibition of this gycosyltransferase may inhibit tumor progression in breast, lung and colon. Further, activation of this gycosyltransferase may inhibit tumor progression in ovary.

As used herein, the term “formyl transferase domain” includes an amino acid sequence of about 20-150 amino acid residues in length and having a bit score for the alignment of the sequence to the formyl transferase domain (HMM) of at least 30. Preferably, a formyl transferase domain includes at least about 40-130 amino acids, more preferably about 60-110 amino acid residues, or about 70- 100 amino acids and has a bit score for the alignment of the sequence to the glycosyltransferase domain (HMM) of at least 50 or greater. The formyl transferase domain (HMM) has been assigned PFAM Accession Number PF00551. The formyl transferase domain (amino acids 119-220 of SEQ ID NO:14) of human 50226 aligns with a consensus amino acid sequence derived from a hidden Markov model.

In a preferred embodiment a 50226 polypeptide or protein has a formyl transferase domain” or a region which includes at least about 20-150 more preferably about 50-125 or 70-100 amino acid residues and has at least about 70% 80% 90% 95%, 99%, or 100% homology with a “formyl transferase domain,” e.g., the formyl transferase domain of human 50226 (e.g., residues 119 to 220 of SEQ ID NO:14).

To identify the presence of a “formyl transferase” domain in a 50226 protein sequence, and make the determination that a polypeptide or protein of interest has a particular profile, the amino acid sequence of the protein can be searched against a database of HMMs

Human 58764

The human 58764 sequence (SEQ ID NO:16), which is approximately 1797 nucleotides long including untranslated regions, contains a predicted methionine-initiated coding sequence of about 972 nucleotides (nucleotides 215-1186 of SEQ ID NO:16; 1-972 of SEQ ID NO:18, not including the terminal codon). The coding sequence encodes a 324 amino acid protein (SEQ ID NO:17). The protein contains a signal sequence from about amino acids 1 to 63. This mature protein form is approximately 261 amino acid residues in length (from about amino acid 64 to amino acid 324 of SEQ ID NO:17).

An alignment of the acyltransferase domain of human 58764 with a consensus amino acid sequence derived from a hidden Markov model (HMM) from PFAM shows the consensus amino acid sequence (SEQ ID NO:46) aligns with amino acids 115 to 300 of SEQ ID NO:17.

A BLAST alignment of human 58764 with a consensus amino acid sequence derived from a ProDomain “CG11757” (PD107349; Release 2001.1) shows amino acid residues 7 to 222 of the 260 amino acid consensus sequence (SEQ ID NO:47) aligns with the “CG11757” domain of human 58764, amino acid residues 91 to 293 of SEQ ID NO:17.

A BLAST alignment of human 58764 with a consensus amino acid sequence derived from a ProDomain “CG11757” (PD260979; Release 2001.1) shows amino acid residues 28 to 50 of the 63 amino acid consensus sequence (SEQ ID NO:48) aligns with the “CG 11757” domain of human 58764, amino acid residues 300 to 322 of SEQ ID NO:17.

Human 58764 contains the following regions or other structural features: one predicted acyltransferase domain (PFAM Accession Number PF01553) located at about amino acid residues 115-300 of SEQ ID NO:17; two or three predicted transmembrane domains (predicted by MEMSAT, Jones et al. (1994) Biochemistry 33:3038-3049) which extend from about amino acid residues 51-74, 124-141 and 159-176 of SEQ ID NO:17; one predicted N-glycosylation site (PS00001) located at about amino acids 5-8 of SEQ ID NO:17; one predicted protein kinase C phosphorylation site (PS00005) located at about amino acids 151-153 of SEQ ID NO:17; two predicted casein kinase II phosphorylation sites (PS00006) located at about amino 98-101 and 289-292 of SEQ ID NO:17; one predicted tyrosine kinase phosphorylation site (PS00007) located at about amino acids 23-261 of SEQ ID NO:17; three predicted N-myristoylation sites (PS00008) located at about amino acids 91-96, 199-204 and 313-318 of SEQ ID NO:17; and five predicted dileucine motifs in the tail located at about amino acids 53-54, 63-64, 168-169, 169-170 and 192-193 of SEQ ID NO:17.

In one embodiment, a 58764 family member can include at least one acyltransferase domain (PFAM Accession Number PF01553); and at least one, and preferably two or three transmembrane domains. Furthermore, a 58764 family member can include at least one N-glycosylation site (PS00001); at least one protein kinase C phosphorylation site (PS00005); at least one, and preferably two casein kinase II phosphorylation sites (PS00006); at least one predicted tyrosine kinase phosphorylation site (PS00007); at least one, two and preferably three N-myristolyation sites (PS00008); at least one, two, three, four and preferably five predicted dileucine motifs in the tail.

A hydropathy plot of human 58764 was performed. Polypeptides of the invention include fragments which include: all or part of a hydrophobic sequence, e.g., the sequence from about amino acid 125 to 140, from about 160 to 180, and from about 225 to 235 SEQ ID NO:17; all or part of a hydrophilic sequence, e.g., the sequence from about amino acid 85 to 90, from about 155 to 125, and from about 240 to 250 of SEQ ID NO:17; a sequence which includes a Cys, or a glycosylation site.

As referred to herein, acyltransferases preferably include a catalytic domain of about 100-250 amino acid residues in length, preferably about 130-200 amino acid residues in length, or more preferably about 160-200 amino acid residues in length. An acyltransferase domain typically includes at least one of four blocks of homology commonly found in members of the acyltransferase family. The four blocks are each characterized by the following motifs: (1) [NX]—H—[RQ]-S—X-[LYIM]-D, SEQ ID NO:49 (2) G-X—[IF]—F—I—[RD]-R. SEQ ID NO:50; (3) F—[PLI]-E-G-[TG]-R—[SX]—[RX], SEQ ID NO:51; and (4) [VI]—[PX]—[IVL]-[IV]—P—[VI], SEQ ID NO:52. Specificity of an acyltransferase for acylation of a particular lipid target can be predicted by the presence of sequences within the four blocks, whereby particular amino acid residues are associated with particular classes of acyltransferases (as described in Lewin et al., (1999) Biochemistry 38:5764-71, for example, the contents of which are incorporated herein by reference). For example, 58764 contains some residues in these blocks of homology that are typically found in LPAATs and not typically found in GPATs. Based on these sequence similarities, the 58764 molecules of the present invention are predicted to have similar biological activities as acyltransferase family members. Thus, the molecules of the present invention may be involved in one or more of: 1) the transfer of an acyl chain to a lipid precursor; 2) the regulation of lipid biosynthesis; 3) the regulation of wound healing; 4) the regulation of platelet aggregation; 5) the modulation of mitogenesis; 6) the modulation of cellular differentiation; 7) the modulation of actin cytoskleleton remodeling; 8) the regulation of monocyte chemotaxis; 9) the modulation of neurite retraction; 10) the modulation of vasoconstriction; 11) the modulation of glutamate and glucose uptake by astrocytes; 12) the modulation of tumor cell growth and invasion; or 13) the formation of synaptic-like microvesicles. Thus, the 58764 molecules can act as novel diagnostic targets and therapeutic agents for controlling acyltransferase-related disorders, for example, such as those diseases associated with the activities described above. As the 58764 molecules have homology to known acyltransferases, they are expected to be involved in controlling similar disorders.

The 26199, 33530, 33949, 47148, 50226, and 58764 proteins contain a significant number of structural characteristics in common with members of the transferase family. The present invention is based, at least in part, on the discovery of novel transferase family members, referred to herein as “transferase” nucleic acid and protein molecules.

A 26199, 33530, 33949, 47148, 50226, or 58764 polypeptide can include a “transferase domain” or regions homologous with an “transferase domain”.

To identify the presence of a “transferase” domain in a 26199, 33530, 33949, 47148, 50226, or 58764 protein sequence, and make the determination that a polypeptide or protein of interest has a particular profile, the amino acid sequence of the protein can be searched against a database of HMMs (e.g., the Pfam database, release 2.1) using the default parameters. For example, the hmmsf program, which is available as part of the HMMER package of search programs, is a family specific default program for MILPAT0063 and a score of 15 is the default threshold score for determining a hit. Alternatively, the threshold score for determining a hit can be lowered (e.g., to 8 bits). A description of the Pfam database can be found in Sonhammer et al., (1997) Proteins 28(3):405-420 and a detailed description of HMMs can be found, for example, in Gribskov et al., (1990) Meth. Enzymol. 183:146-159; Gribskov et al., (1987) Proc. Natl. Acad. Sci. USA 84:4355-4358; Krogh et al., (1994) J. Mol. Biol. 235:1501-1531; and Stultz et al., (1993) Protein Sci. 2:305-314, the contents of which are incorporated herein by reference.

For further identification of domains in a 26199, 33530, 33949, 47148, 50226, or 58764 protein sequence, and make the determination that a polypeptide or protein of interest has a particular profile, the amino acid sequence of the protein can be searched against a database of domains, e.g., the ProDom database (Corpet et al. (1999), Nucl. Acids Res. 27:263-267). The ProDom protein domain database consists of an automatic compilation of homologous domains. Current versions of ProDom are built using recursive PSI-BLAST searches (Altschul S F et al. (1997) Nucleic Acids Res. 25:3389-3402; Gouzy et al. (1999) 23:333-340) of the SWISS-PROT 38 and TREMBL protein databases. The database automatically generates a consensus sequence for each domain. A BLAST search was performed against the HMM database resulting in the identification of a “transferase” domain(s) in the amino acid sequence of human 26199 at about residues 7 to 120, 123 to 226, and 15 to 208 of SEQ ID NO:2 having 44%, 27% and 29% identity over those residues respectively; of human 33530 at about residues 367 to 415, 17 to 95, 280 to 413, 96 to 143, 109 to 155, and 216 to 382 of SEQ ID NO:5 having 91%,49%, 35%, 56%, 40% and 30% identity over those residues respectively; of human 33949 at about residues 1 to 102, 103 to 229, 347 to 406, 568 to 608 and 484 to 521 (two local alignments), and 273 to 346 of SEQ ID NO:8 having 100%, 49%, 64%, 100%, 39%, and 56% identity over those residues respectively; of human 47148 at about residues 1 to 135, 200 to 471, and 549 to 662 of SEQ ID NO:11 having 80%, 37% and 64% identity over those residues respectively; of human 50226 at about residues 117 to 221 and 238 to 355 of SEQ ID NO:14 having 39% and 29% identity over those residues respectively; and of human 58764 at about residues 91 to 293 and 300 to 322 of SEQ ID NO:17 having 41% and 56% identity over those residues respectively.

An additional method to identify the presence of a “transferase” domain in a 26199, 33530, 33949, 47148, 50226, or 58764 protein sequence, and make the determination that a polypeptide or protein of interest has a particular profile, the amino acid sequence of the protein can be searched against a SMART database (Simple Modular Architecture Research Tool) of HMMs as described in Schultz et al. (1998), Proc. Natl. Acad. Sci. USA 95:5857 and Schultz et al. (2000) Nucl. Acids Res 28:231. The database contains domains identified by profiling with the hidden Markov models of the HMMer2 search program (R. Durbin et al. (1998) Biological sequence analysis: probabilistic models of proteins and nucleic acids. Cambridge University Press). The database also is extensively annotated and monitored by experts to enhance accuracy. For example, a search was performed against the HMM database resulting in the identification of a “ricin_—3” domain in the amino acid sequence of human 33949 at about residues 476 to 607 of SEQ ID NO:8.

In one embodiment, 26199, 33530, 33949, 47148, and 58764 proteins include at least one transmembrane domain. As used herein, the term “transmembrane domain” includes an amino acid sequence of about 14 amino acid residues in length that spans a phospholipid membrane. More preferably, a transmembrane domain includes about at least 15, 16, 17, 18, 20, 21, 23 or 24 amino acid residues and spans a phospholipid membrane. Transmembrane domains are rich in hydrophobic residues, and typically have an α-helical structure. In a preferred embodiment, at least 50%, 60%, 70%, 80%, 90%, 95% or more of the amino acids of a transmembrane domain are hydrophobic, e.g., leucines, isoleucines, tyrosines, or tryptophans. Transmembrane domains are described in, for example, Zagotta W. N. et al., (1996) Annual Rev. Neuronsci. 19: 235-63, the contents of which are incorporated herein by reference.

In a preferred embodiment, 26199, 33530, 33949, 47148, and 58764 polypeptides or proteins have at least one transmembrane domain or a region which includes at least 15, 16, 17, 18, 20, 21, 23 or 24 amino acid residues and has at least about 60%, 70% 80% 90% 95%, 99%, or 100% homology with a “transmembrane domain,” e.g., at least one transmembrane domain of human 26199, 33530, 33949, 47148, or 58764 (e.g., amino acid residues 33-49 and 74-94 of SEQ ID NO:2; amino acids 85-105 of SEQ ID NO:5; amino acids 8-28, 150-168, and 268-284 of SEQ ID NO:8; amino acids 106-127 and 168-192 of SEQ ID NO:11; and amino acids 51-74, 124-141, and 159-176 of SEQ ID NO:17).

In another embodiment, a 26199, 33530, 33949, 47148, or 58764 protein includes at least one “non-transmembrane domain.” As used herein, “non-transmembrane domains” are domains that reside outside of the membrane. When referring to plasma membranes, non-transmembrane domains include extracellular domains (i.e., outside of the cell) and intracellular domains (i.e., within the cell). When referring to membrane-bound proteins found in intracellular organelles (e.g., mitochondria, endoplasmic reticulum, peroxisomes and microsomes), non-transmembrane domains include those domains of the protein that reside in the cytosol (i.e., the cytoplasm), the lumen of the organelle, or the matrix or the intermembrane space (the latter two relate specifically to mitochondria organelles). The C-terminal amino acid residue of a non-transmembrane domain is adjacent to an N-terminal amino acid residue of a transmembrane domain in a naturally-occurring 26199, 33530, 33949, 47148, or 58764, or 26199-, 33530-, 33949-, 47148-, or 58764-like protein.

In a preferred embodiment, a 26199, 33530, 33949, 47148, or 58764 polypeptide or protein has a “non-transmembrane domain” or a region which includes at least about 1-150, preferably about 5-140, more preferably about 10-130, and even more preferably about 16-120 amino acid residues, and has at least about 60%, 70% 80% 90% 95%, 99% or 100% homology with a “non-transmembrane domain”, e.g., a non-transmembrane domain of human 26199, 33530, 33949, 47148, or 58764 (e.g., residues 1-32, 50-73 or 95-229 of SEQ ID NO:2; residues 1-84 and 105-416 of SEQ ID NO:5; residues 1-8, 29-149, 169-263, and 285-608 of SEQ ID NO:8; residues 1-105, 128-167 and 193-662 of SEQ ID NO:11; or residues 1-50, 75-123, 142-158, and 177-324 of SEQ ID NO:17). Preferably, a non-transmembrane domain is capable of catalytic activity.

A non-transmembrane domain located at the N-terminus of a 26199, 33530, 33949, 47148, or 58764 protein or polypeptide is referred to herein as an “N-terminal non-transmembrane domain.” As used herein, an “N-terminal non-transmembrane domain” includes an amino acid sequence having about 1-150, preferably about 2-125, more preferably about 4-110, or even more preferably about 7-105 amino acid residues in length and is located outside the boundaries of a membrane. For example, an N-terminal non-transmembrane domain is located at about amino acid residues 1-32 of SEQ ID NO:2.

Similarly, a non-transmembrane domain located at the C-terminus of a 26199, 33530, 33949, 47148, or 58764 protein or polypeptide is referred to herein as a “C-terminal non-transmembrane domain.” As used herein, a “C-terminal non-transmembrane domain” includes an amino acid sequence having about 1-600, preferably about 75-525, preferably about 125-475, more preferably about 134-469 amino acid residues in length and is located outside the boundaries of a membrane. For example, a C-terminal non-transmembrane domain is located at about amino acid residues 95-229 of SEQ ID NO:2.

In another embodiment, a 33949, 50226, or 58764 molecule can further include a signal sequence. As used herein, a “signal sequence” refers to a peptide of about 10-80 amino acid residues in length which occurs at the N-terminus of secretory and integral membrane proteins and which contains a majority of hydrophobic amino acid residues. For example, a signal sequence contains at least about 12-70 amino acid residues, preferably about 15-65 amino acid residues, more preferably about 17-63 amino acid residues, and has at least about 40-70%, preferably about 50-65%, and more preferably about 55-60% hydrophobic amino acid residues (e.g., alanine, valine, leucine, isoleucine, phenylalanine, tyrosine, tryptophan, or proline). Such a “signal sequence”, also referred to in the art as a “signal peptide”, serves to direct a protein containing such a sequence to a lipid bilayer. For example, in one embodiment, a 33949 protein contains a signal sequence of about amino acids 1-37 of SEQ ID NO:8. The “signal sequence” is cleaved during processing of the mature protein. The mature 33949 protein corresponds to amino acids 38-608 of SEQ ID NO:8. The 50226 or 58764 protein may include a signal sequence, and thus the mature 50226 or 58764 protein may correspond to amino acids 18-389 of SEQ ID NO:14 or amino acids 64-324 of SEQ ID NO:17 respectively.

As the 26199, 33530, 33949, 47148, 50226, or 58764 polypeptides of the invention may modulate 26199-, 33530-, 33949-, 47148-, 50226-, or 58764-mediated activities, they may be useful for developing novel diagnostic and therapeutic agents for 26199-, 33530-, 33949-, 47148-, 50226-, or 58764-mediated or related disorders, as described below.

As used herein, a “26199, 33530, 33949, 47148, 50226, or 58764 activity”, “biological activity of 26199, 33530, 33949, 47148, 50226, or 58764” or “functional activity of 26199, 33530, 33949, 47148, 50226, or 58764”, refers to an activity exerted by a 26199, 33530, 33949, 47148, 50226, or 58764 protein, polypeptide or nucleic acid molecule on e.g., a 26199-, 33530-, 33949-, 47148-, 50226-, or 58764-responsive cell or on a 26199, 33530, 33949, 47148, 50226, or 58764 substrate, e.g., a lipid or protein substrate, as determined in vivo or in vitro. In one embodiment, a 26199, 33530, 33949, 47148, 50226, or 58764 activity is a direct activity, such as an association with a 26199, 33530, 33949, 47148, 50226, or 58764 target molecule. A “target molecule” or “binding partner” is a molecule with which a 26199, 33530, 33949, 47148, 50226, or 58764 protein binds or interacts in nature, e.g., a lipid to which the 26199, 33530, 33949, 47148, 50226, or 58764 protein attaches an acyl chain. A 26199, 33530, 33949, 47148, 50226, or 58764 activity can also be an indirect activity, e.g., a cellular signaling activity mediated by interaction of the 26199, 33530, 33949, 47148, 50226, or 58764 protein with a 26199, 33530, 33949, 47148, 50226, or 58764 ligand.

The transferase molecules of the present invention are predicted to modulate and facilitate cell proliferation, differentiation, motility, and apoptosis. Thus, the transferase molecules of the present invention may play a role in cellular growth signaling mechanisms. As used herein, the term “cellular growth signaling mechanism” includes signal transmissions from cell receptors, e.g., growth factor receptors, which regulate one or more of the following: 1) cell transversal through the cell cycle, 2) cell differentiation, 3) cell migration and patterning, and 4) programmed cell death. Throughout development and in the adult organism, cell fate and activity is determined, in part, by extracellular and intracellular stimuli, e.g., growth factors, angiogenic factors, chemotactic factors, neurotrophic factors, cytokines, and hormones. These stimuli act on their target cells by initiating signal transduction cascades that alter the pattern of gene expression and metabolic activity so as to mediate the appropriate cellular response. The transferase molecules of the present invention are predicted to be involved in the initiation or modulation of cellular signal transduction pathways that modulate cell growth, differentiation, migration and/or apoptosis. Thus, the transferase molecules, by participating in cellular growth signaling mechanisms, may modulate cell behavior and act as therapeutic agents for controlling cellular proliferation, differentiation, migration, and apoptosis.

Altered expression of factors (e.g., a transferase molecule) involved in the regulation of signaling pathways associated with cell growth, differentiation, migration, and apoptosis can lead to perturbed cellular proliferation, which in turn can lead to cellular proliferative and/or differentiative disorders. As used herein, a “cellular proliferative disorder” includes a disorder, disease, or condition characterized by a deregulated, e.g., upregulated or downregulated, growth response. As used herein, a “cellular differentiative disorder” includes a disorder, disease, or condition characterized by aberrant cellular differentiation. Thus, the transferase molecules can act as novel diagnostic targets and therapeutic agents for controlling cellular proliferative and/or differentiative disorders.

Examples of cellular proliferative and/or differentiative disorders include cancer, e.g., carcinoma, sarcoma, metastatic disorders or hematopoietic neoplastic disorders, e.g., leukemias. A metastatic tumor can arise from a multitude of primary tumor types, including but not limited to those of prostate, colon, lung, breast and liver origin.

The 26199, 33530, 33949, 47148, 50226, and 58764 nucleic acid and protein of the invention can be used to treat and/or diagnose a variety of proliferative disorders. E.g., such disorders include hematopoietic neoplastic disorders.

Gene Expression Analysis of 26199, 33530, 33949, 50226 and 58764

Human 26199, 33530, 33949, 50226 or 58764 expression was measured by TaqMan® quantitative PCR (Perkin Elmer Applied Biosystems) in cDNA prepared from a variety of normal and diseased (e.g., cancerous) human tissues or cell lines.

26199 was identified as being induced in MCF10A and MCF10AT3B human breast epithelial cells, following stimulation with epidermal growth factor (EGF). Taqman results for 26199 on cDNA from untreated MCF10A, untreated MCF10AT3B cells and cells treated with 10 ng EGF/ml for 0.5, 1, 2, 4 and 8 hours is shown in the following Table 2. In the EGF treated MCF10A and MCF10AT3B panel, 26199 expression increased in MCF10A cells at 1 hour through 4 hours post EGF-treatment. This was consistent with the array data, although slightly delayed, as the array data showed an increase in 26199 expression at 30 minutes post EGF treatment.

The 25K array was profiled with probes generated from untreated MCF10A cells and MCF10A cells treated with 10 ng EGF/ml for 0.5, 1, 2, 4 and 8 hours. The 25K array was also profiled with probes generated from untreated MCF10AT3B cells and MCF10AT3B cells treated with 10 ng EGF/ml for 0.5, 1, 2, 4 and 8 hours. 26199 expression increased following EGF treatment.

The MPGv3.0 array was profiled with probes generated from 4 normal breast tissue samples, 4 ductal carcinoma in situ (DCIS) samples, 4 invasive ductal carcinoma (IDC) samples and 3 invasive lobular carcinoma (ILC) samples. A clone representing 26199 showed expression levels at 2.6-4.5 the median array intensity of the normal breast samples in 3/4 DCIS samples.

This discrepancy may be explained by the fact that RNAs from different EGF-treated MCF10A cell experiments were used for the array and Taqman experiments. MCF10AT3B cells also showed an increase in 26199 expression at 1 hour post EGF-treatment, but overall expression levels were low (Ct values ˜30).

TABLE 2


26199 Expression in EGF-treated MCF10A and MCF10AT3B Cells

MCF10A 0 hr	27.0	15.7	0.4
MCF10A 0.5 hr	27.0	15.6	0.4
MCF10A 1 hr	25.5	15.9	1.3
MCF10A 2 hr	25.2	15.8	1.4
MCF10A 4 hr	25.7	15.9	1.1
MCF10A 8 hr	26.5	16.0	0.7
MCF3B 0 hr	30.7	17.8	0.1
MCF3B 0.5 hr	31.0	17.8	0.1
MCF3B 1 hr	29.1	17.3	0.3
MCF3B 2 hr	30.1	18.4	0.3
MCF3B 4 hr	30.1	18.1	0.2
MCF3B 8 hr	30.3	17.7	0.2

The following Table 3 shows the Taqman results for an oncology panel (Phase I) of human tissues. 26199 expression was upregulated by 4-16-fold in 6/6 breast tumor samples versus 3/4 normal breast samples. Lung tumors uniformly expressed increased levels of 26199 in comparison to normal lung samples.

TABLE 3


26199 Expression in Clinical Tumor Samples

Average	Average	Relative
26199	Beta 2	Expression

Breast N	37.0	22.2	0.0
Breast N	38.5	20.6	0.0
Breast N	33.2	17.2	0.0
Breast N	31.3	19.0	0.6
Breast T	30.8	17.3	0.3
Breast T	31.8	17.9	0.2
Breast T	28.1	16.2	0.8
Breast T	30.0	16.5	0.3
Breast T	32.2	18.2	0.2
Breast T	32.5	19.3	0.4
Ovary N	28.9	17.3	1.1
Ovary N	30.2	18.7	1.1
Ovary N	31.2	19.1	0.8
Ovary N	34.9	22.3	0.5
Ovary T	33.1	18.3	0.1
Ovary T	32.5	17.6	0.1
Ovary T	30.3	16.9	0.3
Ovary T	32.8	17.8	0.1
Ovary T	32.3	17.3	0.1
Ovary T	35.1	19.2	0.0
Ovary T	33.3	20.3	0.4
Ovary T	33.0	16.5	0.0
Lung N	40.0	21.8	0.0
Lung N	40.0	18.5	0.0
Lung N	32.7	16.2	0.0
Lung N	38.0	15.6	0.0
Lung T	29.5	16.1	0.3
Lung T	27.1	16.0	1.4
Lung T	31.8	17.4	0.2
Lung T	31.4	16.5	0.1
Lung T	30.4	18.7	1.0
Lung T	32.3	18.6	0.3
Lung T	30.5	17.2	0.3

The following Table 4 shows the Taqman results for another oncology (Phase II) panel of human tissues. Breast, ovary, colon and lung tumors all expressed 26199. Differential expression between tumor and normal tissues was most significant in colon and lung tissues.

TABLE 4


26199 Expression in Clinical Tumor Samples

Average	Average	Relative
26199	Beta 2	Expression

Colon N	32.2	16.9	0.0
Colon N	35.4	21.1	0.0
Colon N	30.0	18.0	0.3
Colon N	31.2	16.8	0.0
Colon T	31.0	16.2	0.0
Colon T	26.8	17.2	1.3
Colon T	31.5	16.0	0.0
Colon T	30.1	17.0	0.1
Colon T	29.7	16.2	0.1
Colon T	32.4	16.0	0.0
Liver Met	30.7	17.3	0.1
Liver Met	31.3	19.6	0.3
Liver Met	31.6	17.8	0.1
Liver Met	31.9	17.7	0.1
Liver Nor	33.0	16.3	0.0
Liver Nor	36.8	22.6	0.0
Brain N	28.3	19.6	2.5
Brain N	28.8	20.2	2.7
Brain N	30.4	19.5	0.5
Brain N	27.3	19.5	4.5
Astrocytes	33.5	21.1	0.2
Brain T	36.3	16.6	0.0
Brain T	35.6	17.4	0.0
Brain T	34.0	18.2	0.0
Brain T	32.2	17.0	0.0
Brain T	33.6	19.2	0.0
HMVEC	29.9	15.9	0.1
HMVEC	30.0	16.5	0.1
Placenta	36.3	22.1	0.0
Fetal Adren	34.9	23.7	0.4
Fetal Adren	25.4	16.2	1.7
Fetal Liver	27.9	19.7	3.3
Fetal Liver	31.5	18.3	0.1

For Taqman results on the phase I tissue panel, highest expression of 26199 orthologs is found in normal brain cortex as shown in the following Table 5.

TABLE 5


26199 Expression w/β2 in Normal Tissues

Tissue Type	Mean	β 2 Mean	∂∂ Ct	Expression

Artery normal	31.98	21.32	10.66	0.6159
Vein normal	33.19	19.65	13.54	0.084
Aortic SMC EARLY	29.74	20.91	8.82	2.205
Coronary SMC	29.25	21.77	7.48	5.6014
Static HUVEC	26.98	20.16	6.83	8.82
Shear HUVEC	26.61	20.43	6.18	13.7445
Heart normal	26.57	18.23	8.35	3.0648
Heart CHF	26.59	18.58	8.02	3.8658
Kidney	27.37	19.32	8.04	3.7863
Skeletal Muscle	27.2	21.36	5.84	17.4576
Adipose normal	34.56	19.29	15.28	0.0251
Pancreas	28.25	20.68	7.58	5.2444
primary osteoblasts	28.33	18.55	9.79	1.1335
Osteoclasts (diff)	36.87	16.93	19.94	0
Skin normal	30.56	20.45	10.1	0.9112
Spinal cord normal	30	20.06	9.94	1.018
Brain Cortex normal	24.36	20.31	4.04	60.5806
Brain Hypothalamus normal	27.36	20.49	6.88	8.5196
Nerve	29.99	23.6	6.39	11.9239
DRG (Dorsal Root Ganglion)	28.39	20.9	7.5	5.5435
Glial Cells (Astrocytes)	27.25	22.14	5.12	28.8557
Glioblastoma	30.12	17.41	12.71	0.1492
Breast normal	28.45	19.5	8.95	2.022
Breast tumor	26.3	17.65	8.64	2.498
Ovary normal	27.08	19.48	7.6	5.1543
Ovary Tumor	28.14	19.61	8.52	2.7241
Prostate Normal	28.88	18.95	9.93	1.0287
Prostate Tumor	27.23	17.13	10.11	0.908
Epithelial Cells (Prostate)	27.35	21.06	6.29	12.7797
Colon normal	30.91	17.58	13.34	0.0968
Colon Tumor	26.05	18.32	7.73	4.7102
Lung normal	33.2	17.39	15.81	0.0174
Lung tumor	26.78	18.11	8.66	2.4636
Lung COPD	31.59	18.16	13.43	0.0906
Colon IBD	34.27	17.11	17.16	0.0068
Liver normal	32.2	19.23	12.97	0.1251
Liver fibrosis	29.07	20.81	8.27	3.2508
Dermal Cells- fibroblasts	31.31	19.29	12.02	0.2408
Spleen normal	27.47	18.92	8.55	2.668
Tonsil normal	28.11	16.62	11.49	0.3465
Lymph node	28.27	18.01	10.26	0.8155
Small Intestine	31.52	19.39	12.14	0.2223
Skin-Decubitus	29.75	19.63	10.12	0.8986
Synovium	32.38	18.95	13.44	0.09
BM-MNC (Bone marrow	26.89	16.43	10.46	0.7124
mononuclear cells)
Activated PBMC	30.66	15.45	15.21	0.0264

The following Table 6 shows the Taqman results of an oncology cell lines panel. 26199 is expressed in many tumor cell lines. MCF-7 human breast cancer cells is expressed at the highest levels.

TABLE 6


26199 Expression in Cell Lines

Average	Average	Relative
26199	B-2	Expression

MCF-7	24.5	19.0	45.9
ZR75	26.6	18.7	8.6
T47D	25.5	18.4	15.6
MDA 231	26.8	17.3	2.8
MDA 435	27.5	16.3	0.9
DLD-1	25.0	19.4	42.5
SW 480	27.5	16.9	1.4
SW 620	25.7	18.6	15.0
HCT 116	26.5	18.4	7.7
HT 29	27.7	16.1	0.7
Colo 205	25.8	15.3	1.4
NCIH 125	26.6	17.9	5.3
NCIH 67	25.6	18.8	18.8
NCIH 322	27.0	18.6	6.1
NCIH 460	26.6	17.2	3.2
A549	25.9	18.6	14.0
NHBE	27.7	19.0	5.0

Confirming previous Taqman results, in a breast cancer cell model panel, 26199 showed increased expression in MCF10A cells treated with EGF and high expression in MCF-7 cells as shown in the following Table 7.

TABLE 7


26199 Expression in Breast Cancer Cell Model Panel

Tissue Type	26199.2Mean	β 2 Mean	∂∂ Ct	Expression

MCF10MS	33.07	20.2	12.87	0.13
MCF10A	28.86	19.93	8.93	2.05
MCF10AT.cl1	27.27	20	7.27	6.48
MCF10AT.cl3	26.41	19.45	6.96	8.00
MCF10AT1	28.55	20.47	8.08	3.70
MCF10AT3B	28.27	20.23	8.04	3.79
MCF10CA1a.cl1	33.88	17.32	16.57	0.01
MCF10CA1a.cl1	33.52	24.69	8.83	2.20
Agar
MCF10A.m25	31.13	24.23	6.9	8.37
Plastic
MCF10CA Agar	30.59	22.17	8.41	2.93
MCF10CA Plastic	29.75	21.52	8.22	3.35
MCF3B Agar	29.02	22.31	6.71	9.59
MCF3B Plastic	28.29	22.19	6.1	14.58
MCF10A EGF 0 hr	26.32	17.72	8.61	2.57
MCF10A EGF 0.5 hr	25.47	17.66	7.8	4.47
MCF10A EGF 1 hr	25.24	17.77	7.47	5.62
MCF10A EGF 2 hr	24.84	17.93	6.9	8.37
MCF10A EGF 4 hr	25.77	17.58	8.19	3.42
MCF10A EGF 8 hr	25.7	18.02	7.68	4.88
MCF10A IGF1A 0 hr	28.02	21.95	6.07	14.94
MCF10A IGF1A	28.84	22.38	6.46	11.32
0.5 hr
MCF10A IGF1A 1 hr	28.61	21.93	6.68	9.75
MCF10A IGF1A 3 hr	28.55	21.86	6.7	9.65
MCF10A IGF1A 24 hr	27.09	21.53	5.56	21.20
MCF10AT3B.cl5	28.56	22.27	6.29	12.78
Plastic
MCF10AT3B.cl6	29.16	21.9	7.25	6.55
Plastic
MCF10AT3B.cl3	29.12	21.88	7.25	6.59
Plastic
MCF10AT3B.cl1	28.68	22.09	6.59	10.34
Plastic
MCF10AT3B.cl4	28.85	21.75	7.09	7.31
Plastic
MCF10AT3B.cl2	28.84	22.13	6.71	9.55
Plastic
MCF10AT3B.cl5	31.66	24.07	7.6	5.15
Agar
MCF10AT3B.cl6	30.9	24.27	6.63	10.13
Agar
MCF-7	27.43	23.34	4.09	58.52
ZR-75	28.2	21.51	6.7	9.65
T47D	28.65	21.72	6.93	8.20
MDA-231	29.11	20.47	8.65	2.49
MDA-435	32.42	20.43	11.99	0.25
SkBr3	28.41	20.93	7.47	5.62
Hs578Bst	30.59	19.98	10.61	0.64
Hs578T	28.68	19.93	8.74	2.33
MCF10AT3B Agar	31.95	26.23	5.71	19.04

For Taqman results in the angiogenesis panel, highest expression of 26199 orthologs is found in Wilm's tumor as shown in the following Table 8.

TABLE 8


Expression of 26199 w/β2 in the Angiogenesis Panel

Tissue Type	26199.2Mean	β 2 Mean	∂∂ Ct	Expression

ONC 101 Hemangioma	29.19	20.93	8.26	3.26
ONC 102 Hemangioma	28.2	19.48	8.72	2.37
ONC 103 Hemangioma	31.97	20.1	11.87	0.27
CHT 1273 Glioblastoma	25.92	20.98	4.95	32.46
CHT 216 Glioblastoma	28.43	18.61	9.82	1.10
CHT 501 Glioblastoma	28.2	21.54	6.66	9.89
NDR 203 Normal Kidney	28.88	21.8	7.08	7.39
PIT 213 Renal Cell Carcinoma	34.1	21.15	12.96	0.13
CHT 732 Wilms Tumor	25.18	20.18	5	31.25
CHT 765 Wilms Tumor	27.55	22.97	4.59	41.67
NDR 295 Skin	32.78	22.2	10.58	0.65
CHT 1424 Uterine Adenocarcinoma	27.2	19.95	7.25	6.55
CHT 1238 Neuroblastoma	28.02	20.8	7.22	6.68
BWH 78 Fetal Adrenal	25.25	19.23	6.02	15.41
BWH 74 Fetal Kidney	26.73	21.11	5.62	20.33
BWH 4 Fetal Heart	27.56	21.14	6.42	11.64
MPI 849 Normal Heart	28.16	20.2	7.96	4.00
CLN 746 Spinal cord	29.54	21.25	8.29	3.21

The following Table 9 shows the Taqman results for an oncology panel (Plate I) of human tissues. 33530 expression was upregulated by 3/8 breast tumor samples versus normal breast samples. 33530 expression was upregulated by 5/7 lung tumor samples versus normal lung samples. 33530 expression is found in both ovary tumors and normal ovary samples.

TABLE 9


33530 Expression in Oncology Plate I

Average	Average	Relative
33530	Beta 2	Expression

Brst N	29.4	22.4	7.8
Brst N	28.9	21.2	4.8
Brst N	25.8	17.5	3.1
Brst N	26.3	19.9	11.4
Brst T	24.0	16.5	5.5
Brst T	30.0	24.1	16.7
Brst T	23.2	15.7	5.5
Brst T	31.0	25.0	16.2
Brst T	25.0	16.0	2.0
Brst T	25.5	16.5	1.9
Brst T	28.3	18.2	0.9
Brst T	26.9	19.5	6.1
Ovry N	25.4	17.4	3.9
Ovry N	28.6	18.4	0.8
Ovry T	28.4	18.3	0.9
Ovry T	27.2	17.6	1.3
Ovry T	26.6	16.8	1.1
Ovry T	28.4	17.6	0.6
Ovry T	27.9	17.3	0.7
Ovry T	30.1	19.2	0.5
Lung N	34.2	22.3	0.3
Lung N	31.2	18.9	0.2
Lung N	25.4	15.0	0.7
Lung N	28.2	16.2	0.2
Lung T	24.3	16.1	3.3
Lung T	25.9	17.0	2.0
Lung T	26.7	17.5	1.7
Lung T	27.4	16.6	0.6
Lung T	26.6	18.8	4.5
Lung T	25.9	17.2	2.4
Lung T	26.7	17.3	1.5

The following Table 10 shows the Taqman results for an oncology panel (Plate II) of human tissues. 33530 expression is found in both colon tumors and normal colon samples as well as normal liver and liver metastases. 33530 expression was downregulated by 6/6 glioblastoma samples versus normal brain samples.

TABLE 10


33530 Expression in Oncology Plate II

Average	Average	Relative
33530	Beta 2	Expression

Colon N	25.6	16.5	1.9
Colon N	28.8	20.7	3.6
Colon N	25.9	18.1	4.4
Colon N	25.3	16.1	1.7
Colon T	23.2	15.7	5.5
Colon T	26.8	16.5	0.8
Colon T	24.0	15.7	3.1
Colon T	25.0	16.3	2.5
Colon T	24.5	16.3	3.4
Colon T	30.4	23.2	6.9
Colon T	25.3	15.7	1.3
Liver Met	26.7	16.9	1.1
Liver Met	28.5	19.3	1.7
Liver Met	27.7	17.5	0.9
Liver Met	28.2	17.3	0.5
Liver Norm	27.4	17.1	0.8
Liver Norm	30.7	22.5	3.6
Brain N	27.5	19.1	3.0
Brain N	26.7	18.6	3.7
Brain N	28.5	19.5	1.9
Brain N	28.0	19.5	2.7
GLIO	29.0	17.8	0.4
GLIO	28.0	16.5	0.3
GLIO	27.8	17.2	0.7
GLIO	27.9	17.1	0.6
GLIO	27.5	16.8	0.6
GLIO	29.3	18.9	0.7
HMVEC	24.9	15.9	1.9
HMVEC	24.4	16.6	4.4
Placenta	25.5	16.0	1.5
Fetal Adrenal	30.8	23.6	6.9
Fetal Adrenal	30.4	23.0	5.9
Fetal Liver	25.8	19.1	10.0
Fetal Liver	27.6	19.3	3.2

The following Table 11 shows the Taqman results of an oncology cell lines panel. 33530 is expressed in many tumor cell lines. NCIH67 cancer cells are expressed at the highest levels.

TABLE 11


33530 Expression in Xenograph Cell Lines

Average	Average	Relative
33530	B-2	Expression

MCF-7	24.5	19.3	27.5
ZR75	24.9	18.0	8.5
T47D	24.9	18.3	10.0
MDA 231	25.5	17.8	4.8
MDA 435	24.4	16.2	3.4
DLD-1	24.1	19.0	29.9
SW 480	24.4	16.6	4.4
SW 620	24.7	18.2	11.1
HCT 116	24.7	18.2	11.0
HT 29	23.5	15.5	3.8
Colo 205	22.2	14.6	4.9
NCIH 125	24.9	17.2	4.9
NCIH 67	22.8	18.6	54.8
NCIH 322	25.4	18.1	6.4
NCIH 460	25.3	17.4	4.1
A549	24.6	19.1	22.0
NHBE	24.9	18.5	11.4

For Taqman results on the phase I tissue panel, highest expression of 33530 orthologs is found in epithelial cells, glial cells and pancreas as shown in the following Table 12.

TABLE 12


Phase 1.2.2 Expression of 33530 w/β2

Tissue Type	Mean	β2 Mean	∂∂ Ct	Expression

Aorta/normal	37.40	24.33	13.07	0.00
Fetal heart/normal	26.36	20.76	5.60	20.69
Heart normal	28.15	19.80	8.36	3.05
Heart/CHF	28.75	21.82	6.93	8.23
Vein/Normal	31.44	20.27	11.17	0.43
Spinal cord/Normal	29.60	19.98	9.62	1.27
Brain cortex/Normal	27.74	21.97	5.77	18.39
Brain hypothalamus/Normal	27.36	21.03	6.33	12.43
Glial cells (Astrocytes)	26.81	22.55	4.26	52.19
Brain/Glioblastoma	27.58	19.24	8.34	3.09
Breast/Normal	30.40	20.10	10.31	0.79
Breast tumor/IDC	27.78	19.53	8.26	3.27
OVARY/Normal	29.29	21.91	7.38	6.00
OVARY/Tumor	29.68	20.37	9.32	1.57
Pancreas	29.50	25.16	4.34	49.38
Prostate/Normal	28.56	20.13	8.44	2.89
Prostate/Tumor	26.51	18.99	7.52	5.45
Colon/normal	31.80	18.59	13.21	0.11
Colon/tumor	25.95	19.55	6.40	11.88
Colon/IBD	29.96	19.32	10.64	0.63
Kidney/normal	28.40	21.43	6.97	7.98
Liver/normal	28.56	19.72	8.84	2.18
Liver fibrosis	28.54	20.98	7.56	5.30
Fetal Liver/normal	27.48	22.30	5.18	27.58
Lung/normal	30.84	18.76	12.08	0.23
Lung/tumor	28.44	19.05	9.39	1.49
Lung/COPD	27.78	19.06	8.73	2.36
Spleen/normal	32.91	21.51	11.40	0.37
Tonsil/normal	30.34	19.01	11.33	0.39
Lymph node/normal	30.65	19.50	11.15	0.44
Thymus/normal	28.21	20.28	7.93	4.11
Epithelial Cells (prostate)	24.93	21.34	3.60	82.76
Endothelial Cells (aortic)	29.20	21.77	7.43	5.80
Skeletal Muscle/Normal	30.83	21.49	9.35	1.54
Fibroblasts (Dermal)	27.80	19.85	7.95	4.06
Skin/normal	30.89	22.13	8.76	2.31
Adipose/Normal	30.81	19.69	11.12	0.45
Osteoblasts (primary)	29.02	21.13	7.89	4.23
Osteoblasts (Undiff)	26.95	19.97	6.98	7.92
Osteoblasts (Diff)	26.73	19.10	7.63	5.07
Osteoclasts	30.45	18.45	12.01	0.24
Aortic SMC Early	26.91	21.36	5.55	21.42
Aortic SMC Late	28.93	24.16	4.78	36.52
shear HUVEC	26.17	21.42	4.76	37.03
static HUVEC	27.71	21.97	5.75	18.65
Osteoclasts (Undiff)	32.78	17.43	15.35	0.02

Table 13 below shows Taqman results for an oncology cell lines panel. 33949 was expressed at high levels in many tumor cell lines, including DLD-1, ZR-75, SW620, NC1H1125 and MCF-7.

TABLE 13


33949 Expression in Xenograph Panel

Average	Average		Relative
33949	B-2	DCt	Expression

MCF-7	22.0	23.1	−1.1	2166.0
ZR75	21.2	23.1	−1.9	3823.8
T47D	21.6	22.0	−0.5	1375.5
MDA 231	21.6	21.0	0.7	619.9
MDA 435	21.8	19.7	2.1	238.2
DLD-1	21.3	25.0	−3.7	12996.0
SW 480	21.7	19.4	2.3	203.1
SW 620	20.9	22.3	−1.4	2657.4
HCT 116	22.1	22.1	0.0	993.1
HT 29	22.2	19.6	2.6	162.1
Colo 205	22.3	18.7	3.6	84.2
NCIH 125	21.1	21.7	−0.6	1489.7
NCIH 67	21.0	22.3	−1.2	2329.5
NCIH 322	22.6	22.2	0.4	737.1
NCIH 460	20.9	21.6	−0.7	1613.3
A549	22.2	23.1	−0.9	1859.6
NHBE	23.2	23.0	0.2	858.6
SKOV-3	24.4	20.8	3.6	82.2
OVCAR-3	24.4	23.9	0.4	732.0
293	22.9	23.9	−1.0	2013.9
293T	22.9	25.3	−2.4	5259.8

In an oncology phase I panel of human tissues, Taqman results showed highest expression of 33949 in the normal brain cortex and kidney pools as shown in the following Table 14. Breast, colon and lung tumor pools expressed higher levels of 33949 than their respective normal tissue counterparts.

TABLE 14


Phase 1.3.3 Expression of 33949

Tissue Type	Mean	β 2 Mean	∂∂ Ct	Expression

Artery normal	31.23	22.2	9.03	1.91
Vein normal	32.41	20.47	11.95	0.25
Aortic SMC EARLY	28.88	21.97	6.91	8.34
Coronary SMC	28.32	23.06	5.26	26.01
Static HUVEC	25.88	20.7	5.18	27.49
Shear HUVEC	26.43	21.16	5.26	26.01
Heart normal	27.95	19.02	8.92	2.06
Heart CHF	26.23	19.27	6.96	8.06
Kidney	24.45	20.72	3.73	75.36
Skeletal Muscle	28.6	21.41	7.18	6.87
Adipose normal	31.28	19.6	11.68	0.30
Pancreas	26.29	21.61	4.67	39.15
primary osteoblasts	27.75	19.37	8.38	3.00
Osteoclasts (diff)	40	17.53	22.47	0.00
Skin normal	29.98	21.37	8.62	2.55
Spinal cord normal	29.07	19.88	9.2	1.71
Brain Cortex normal	23.39	21.23	2.15	225.31
Brain Hypothalamus normal	26.27	21.24	5.03	30.71
Nerve	33.73	23.85	9.88	1.06
DRG (Dorsal Root Ganglion)	27.38	21.94	5.43	23.12
Glial Cells (Astrocytes)	27.7	22.29	5.41	23.60
Glioblastoma	25.72	18.29	7.43	5.78
Breast normal	28.07	20.45	7.62	5.10
Breast tumor	24.44	18.28	6.16	13.98
Ovary normal	25.82	20.36	5.45	22.88
Ovary Tumor	31.1	20.29	10.81	0.56
Prostate Normal	26.41	19.45	6.95	8.09
Prostate Tumor	24.09	17.93	6.16	13.98
Epithelial Cells (Prostate)	26.16	21.29	4.87	34.20
Colon normal	31.93	18.18	13.76	0.07
Colon Tumor	25.5	19.03	6.46	11.32
Lung normal	33.51	18.43	15.07	0.03
Lung tumor	25.87	18.64	7.24	6.64
Lung COPD	28.28	18.41	9.87	1.07
Colon IBD	35.7	18.15	17.56	0.00
Liver normal	32.31	20.04	12.27	0.20
Liver fibrosis	30.41	21.65	8.77	2.30
Dermal Cells- fibroblasts	29.56	20.88	8.69	2.43
Spleen normal	31.32	19.43	11.9	0.26
Tonsil normal	28.16	17.18	10.98	0.49
Lymph node	30.18	18.55	11.64	0.31
Small Intestine	32.89	19.52	13.37	0.09
Skin-Decubitus	29.54	20.52	9.02	1.93
Synovium	33.65	20.19	13.46	0.09
BM-MNC (Bone marrow	31.97	16.69	15.28	0.03
mononuclear cells)
Activated PBMC	32.12	15.93	16.18	0.01

Table 15 below shows Taqman results for oncology phase II panels of human tissues. 33949 was expressed at high levels in a subset of breast, lung, colon, ovarian and brain tumors. Differential expression between tumors and respective normal tissues was most significant in lung tissue. Confirming phase I expression patterns, normal brain tissue expressed very high levels of 33949.

TABLE 15


33949 Expression in Oncology Plate

Tissue Type	Mean	β 2 Mean	∂∂ Ct	Expression

PIT 400 Breast N	24.57	18.06	6.28	12.8686
PIT 271 Breast N	30.05	24.81	5.01	31.0341
PIT 56 Breast N	29.92	21.97	7.71	4.7594
MDA 106 Breast T	26.18	20.25	5.68	19.4377
MDA 234 Breast T	26.1	17.11	8.76	2.3146
NDR 57 Breast T	25.75	18.3	7.21	6.7542
MDA 304 Breast T	28.6	18.4	9.96	1.0005
NDR 58 Breast T	23.85	17.22	6.4	11.8415
NDR 132 Breast T	24.95	20.3	4.42	46.5524
NDR 07 Breast T	29.02	18.8	9.98	0.9868
NDR 12 Breast T	26.93	20.73	5.96	16.0087
PIT 208 Ovary N	23.66	18.22	5.21	27.1106
CHT 620 Ovary N	25.61	19.01	6.37	12.0904
CHT 619 Ovary N	24.16	19.19	4.74	37.4212
CLN 03 Ovary T	26.27	19.18	6.84	8.6986
CLN 05 Ovary T	26.7	18.09	8.37	3.0331
CLN 17 Ovary T	24.8	19.27	5.29	25.471
CLN 07 Ovary T	27.07	18.56	8.28	3.2283
CLN 08 Ovary T	27.11	18.23	8.64	2.498
MDA 216 Ovary T	29.27	20.16	8.88	2.1299
CLN 012 Ovary T	24.94	20.36	4.34	49.3776
MDA 25 Ovary T	25.53	20.97	4.33	49.721
MDA 183 Lung N	28.27	17.23	10.81	0.557
CLN 930 Lung N	29.38	20.37	8.78	2.2828
MDA 185 Lung N	28.19	19.45	8.5	2.7621
MDI 215 Lung T	24.5	18.5	5.76	18.3892
MDA 259 Lung T	23.52	18.9	4.39	47.6956
CHT 832 Lung T	23.47	18.11	5.12	28.8557
MDA 253 Lung T	26.18	17.5	8.46	2.8496
CHT 814 Lung T	23.95	16.45	7.27	6.4791
CHT 911 Lung T	24.47	18.52	5.72	18.9718
CHT 726 Lung T	24.97	16.82	7.92	4.1433
MDA 253 Lung T	24.09	19.9	3.96	64.0348
CHT 845 Lung T	23.81	19.38	4.2	54.5983
NHBE	38.42	18.29	19.9	0
CHT 396 Colon N	32	18.5	13.27	0.10
CHT 519 Colon N	35.76	19.97	15.56	0.00
CHT 416 Colon N	29.23	19.16	9.83	1.10
CHT 452 Colon N	34.25	17.43	16.59	0.01
CHT 398 Colon T	25.08	19.49	5.36	24.43
CHT 805 Colon T	26.63	18.18	8.21	3.38
CHT 528 Colon T	25.23	18.18	6.83	8.82
CHT 368 Colon T	27.58	17.29	10.06	0.94
CHT 372 Colon T	28.23	19.59	8.4	2.95
CHT 01 Liver Met	32.19	18.34	13.61	0.08
CHT 896 Liver Met	28.47	19.28	8.95	2.02
NDR 217 Liver Met	30.55	18.48	11.82	0.28
PIT 260 Liver N	34.7	17.28	17.19	0.01
PIT 229 Liver N	29.16	24.06	4.87	34.20
MGH 16 Brain N	27.71	23.88	3.6	82.76
MCL 53 Brain N	25.63	23.82	1.58	335.64*
MCL 377 Brain N	28.52	24.89	3.4	94.73
MCL 390 Brain N	25.23	23.19	1.8	287.17*
MPI 665 Astrocytes	24.72	19.84	4.65	39.83
CHT 201 Brain T	35.55	20.35	14.97	0.00
CHT 216 Brain T	24.07	17.25	6.58	10.45
CHT 501 Brain T	26.57	20.53	5.8	17.89
CHT 1273 Brain T	24.31	21.39	2.69	155.50*
CHT 828 Brian T	30.73	21.98	8.52	2.72
A24 HMVEC-Arr	25.1	18.09	6.78	9.13
C48 HMVEC-Prol	25.8	20.19	5.38	24.10
CHT 50 Placenta	28.98	24.77	3.98	63.37
BWH 75 Fetal Liver	26.2	19.39	6.58	10.49
BWH 54 Fetal Liver	27.23	21.57	5.42	23.36
PIT 213 Rnal Tumor	36.71	24.85	11.63	0.00
CHT 1424 Endometrial AC	30.24	23.34	6.66	9.89
BWH 58 Fetal Adrenal	32.01	26.05	5.73	18.84
PIT 251 Fetal Adrenal	32.1	26.06	5.81	17.82

Table 16 below shows Taqman results for an MCF10 variants cell model panel (breast cancer cell model panel). Confirming transcription profiling data, 33949 was expressed at highest levels in MCF10AT3B cells grown on agar vs. plastic. 33949 was first identified by transcription profiling as being expressed at higher levels in MCF10AT3B cells grown anchorage-independently vs. anchorage-dependently.

In microarray expression experiments, a human MPGv3.0 array was hybridized with probes generated from various MCF10 cells:

MCF10A (10A)—normal human breast epithelial cell, nontransformed, nontumorigenic

MCF10A.m25 (10A.m25)—clone of MCF10A

MCF10AT.c11 (CL.1)—activated-ras-expressing nontumorigenic clone

MCF10AT.c13 (CL.3)—activated-ras-expressing nontumorigenic clone

MCF10AT1 (AT1)—activated-ras-expressing tumorigenic line, derived from passage of

MCF10AT cells through immunocompromised mice

MCF10AT3B (3B)—activated ras-expressing tumorigenic line, derived from two additional serial passages of MCF10AT1 through immunocompromised mice

Cells were cultured anchorage-dependently on plastic unless indicated by the word ‘agar’ in which case the cells were cultured anchorage-independently in soft agar. 33949 was expressed at the highest levels in MCF10AT3B cells grown anchorage-independently vs. anchorage-dependently.

This result was repeated in independently prepared MCF10AT3B plastic and agar samples as shown in the Taqman results below.

TABLE 16


33949 Expression in MCF10A Variant Cells

Average	Average		Relative
33949	Beta 2	D Ct	Expression

MCF10A -NT	23.7	17.7	6.0	15.7
MCF10AT.c11 -NT	23.1	17.3	5.8	18.5
MCF10AT.c13 -NT	23.4	17.5	6.0	15.8
MCF10MS -NT	23.8	17.5	6.3	12.4
MCF10CA1a.c11 -T	22.3	15.1	7.3	6.4
MCF10AT1 -T	23.9	18.0	5.9	17.0
MCF10AT3B -T	24.1	18.0	6.2	13.8
MCF10AT3B-agar	27.3	23.7	3.6	84.5
MCF10CA1a.cl1-agar	27.1	22.0	5.1	30.0
MCF10A-m25-plastic	27.7	22.1	5.6	21.0
MCF CA-agar	24.0	18.9	5.1	28.9
MCF CA-plastic	23.5	18.3	5.2	27.2
MCF 3B-agar	24.2	20.0	4.2	53.1
MCF 3B-plastic	24.8	19.4	5.4	23.2

The following Table 17 depicts a Taq Man array of 50226 RNA expression relative to the progression of cells through the cell cycle of human colon cancer cells, HCT166 and HCT116 Noc Cells.

TABLE 17


50226 Expression in HCT166 and HCT116 Noc Cells

Average	Average		Relative
50226.1	Beta 2	D Ct	Expression

HCT116 t = 0	23.92	20.28	3.64	80.21
HCT116 t = 3	20.82	17.08	3.74	74.84
HCT116 t = 6	21.26	17.42	3.84	69.83
HCT116 t = 9	21.57	17.78	3.79	72.29
HCT116 t = 12	20.46	16.37	4.09	58.72
HCT116 t = 15	29.10	21.57	7.53	5.43
HCT116 t = 18	21.19	17.26	3.93	65.61
HCT116 t = 21	21.09	17.60	3.49	89.00
HCT116 t = 24	21.27	17.41	3.87	68.63
HCT116 NOC t = 0	23.64	21.51	2.13	228.46
HCT116 Noc t = 3	24.25	22.02	2.23	213.16
HCT116 Noc t = 6	24.07	21.39	2.68	156.04
HCT116 Noc t = 9	23.60	20.77	2.83	140.63
HCT116 Noc t = 15	24.51	22.45	2.06	240.65
HCT116 Noc t = 18	23.95	21.32	2.63	161.54
HCT116 Noc t = 21	24.09	20.99	3.10	116.63
HCT116 Noc t = 24	24.51	21.69	2.83	141.12

For Taqman results on the phase I tissue panel, highest expression of 50226 orthologs is found in normal brain cortex as shown in the following Table 18. Breast, prostate, colon and lung tumor pools expressed higher levels of 50226 than their respective normal tissue counterparts. Normal ovary and prostate pools expressed higher levels of 50226 than their respective tumor tissue counterparts.

TABLE 18


Phase 1.4.3 Expression of 50226.1

Tissue Type	Mean	β 2 Mean	∂∂ Ct	Expression

Artery normal	30.18	23.23	6.95	8.088
Vein normal	31.3	21.28	10.02	0.9665
Aortic SMC EARLY	29.36	22.54	6.82	8.8507
Coronary SMC	29.88	23.15	6.73	9.4204
Static HUVEC	27.09	21.57	5.53	21.7175
Shear HUVEC	27.27	22.45	4.82	35.4026
Heart normal	26.66	19.45	7.21	6.7542
Heart CHF	26.14	19.93	6.21	13.5553
Kidney	28	20.84	7.16	7.0167
Skeletal Muscle	29.7	23.25	6.46	11.3986
Adipose normal	32.58	23.22	9.36	1.5271
Pancreas	30.8	23.16	7.63	5.0483
primary osteoblasts	27.09	20.11	6.98	7.9216
Osteoclasts (diff)	29.05	18.11	10.93	0.5126
Skin normal	31.93	23.18	8.76	2.3146
Spinal cord normal	31.09	21.81	9.28	1.6142
Brain Cortex normal	27.18	23.11	4.07	59.7466
Brain Hypothalamus normal	29.58	22.89	6.7	9.6517
Nerve	30.18	22.61	7.57	5.2626
DRG (Dorsal Root Ganglion)	29.32	22.54	6.78	9.1311
Resting PBMC	28.56	16.98	11.59	0.3255
Glioblastoma	28.11	19.28	8.84	2.1898
Breast normal	31.39	23.15	8.23	3.3191
Breast tumor	27.75	20	7.75	4.6615
Ovary normal	27.11	20.98	6.12	14.3779
Ovary Tumor	29.52	20.84	8.67	2.4551
Prostate Normal	28.54	21.18	7.36	6.1084
Prostate Tumor	28.86	21.33	7.54	5.3919
Colon normal	29.94	20.62	9.32	1.5646
Colon Tumor	25.78	19.79	5.99	15.7337
Lung normal	28.98	19.22	9.77	1.1453
Lung tumor	27	19.61	7.4	5.9208
Lung COPD	30.52	20.83	9.69	1.2107
Colon IBD	31.5	19.93	11.57	0.3278
Liver normal	30.23	21.82	8.41	2.9298
Liver fibrosis	29.7	22.08	7.62	5.0834
Dermal Cells- fibroblasts	27.83	19.76	8.07	3.7212
Spleen normal	30.41	20.82	9.59	1.302
Tonsil normal	28.18	18.33	9.85	1.0836
Lymph node	30.59	20.27	10.32	0.7796
Skin-Decubitus	29.86	21.06	8.8	2.2358
Synovium	30.67	20.34	10.34	0.7742
BM-MNC (Bone marrow	26.86	17.29	9.57	1.3111
mononuclear cells)
Activated PBMC	30.02	19.17	10.85	0.5437
Epithelial Cells (Prostate)	30.75	26.5	4.25	52.556
small Intestine	32.32	24.07	8.25	3.2848

The Taqman results in the following Table 19 also show highest expression of 50226 orthologs in normal brain cortex.

TABLE 19


Phase 1.3.3 Expression of 50226 w/β2

Tissue Type	Mean	β 2 Mean	∂∂ Ct	Expression

Artery normal	30.93	21.29	9.65	1.2447
Vein normal	30.26	19.26	11	0.4883
Aortic SMC EARLY	25.3	19.11	6.2	13.6496
Coronary SMC	27.07	21.04	6.03	15.3566
Static HUVEC	25.16	19.84	5.31	25.2076
Shear HUVEC	24.94	19.89	5.05	30.0811
Heart normal	25.16	18.14	7.02	7.7049
Heart CHF	24.55	18.68	5.87	17.0983
Kidney	26.04	19.78	6.26	13.0482
Skeletal Muscle	27.34	20.53	6.82	8.8814
Adipose normal	29.99	18.93	11.07	0.4668
Pancreas	28.31	20.91	7.4	5.9208
primary osteoblasts	25.76	18.66	7.11	7.2641
Osteoclasts (diff)	32.91	17.1	15.81	0.0174
Skin normal	29.99	20.32	9.66	1.2318
Spinal cord normal	28.36	19.06	9.29	1.5919
Brain Cortex normal	24.93	20.15	4.79	36.272
Brain Hypothalamus normal	27.38	20.47	6.92	8.2866
Nerve	31.3	23.34	7.96	4.03
DRG (Dorsal Root Ganglion)	28.2	21.32	6.87	8.5492
Glial Cells (Astrocytes)	27.02	21.56	5.46	22.6397
Glioblastoma	27.93	17.18	10.74	0.5827
Breast normal	30.11	19.61	10.51	0.6881
Breast tumor	26.22	17.66	8.56	2.6405
Ovary normal	25.93	19.54	6.39	11.9239
Ovary Tumor	30.86	19.39	11.48	0.3501
Prostate Normal	28.35	18.9	9.46	1.4248
Prostate Tumor	25.82	17.18	8.64	2.498
Epithelial Cells (Prostate)	25.31	20.23	5.08	29.6669
Colon normal	28.63	17.5	11.14	0.4447
Colon Tumor	24.52	18.2	6.32	12.5602
Lung normal	32.88	18	14.88	0.0332
Lung tumor	26.56	17.91	8.65	2.4894
Lung COPD	30.25	17.91	12.35	0.1915
Colon IBD	34.87	17.56	17.31	0.0062
Liver normal	28.68	19.39	9.29	1.5919
Liver fibrosis	28.52	21.29	7.24	6.6382
Dermal Cells- fibroblasts	26.73	18.81	7.92	4.1147
Spleen normal	31.04	18.86	12.18	0.2163
Tonsil normal	27.56	16.65	10.91	0.5197
Lymph node	29.31	18.07	11.23	0.4149
Small intestine	29.88	19.14	10.74	0.5847
Skin-Decubitus	29.5	19.95	9.55	1.334
Synovium	37.22	20.08	17.14	0
BM-MNC (Bone marrow	26.81	16.31	10.49	0.6929
mononuclear cells)
Activated PBMC	28.93	15.64	13.29	0.0998

The following Table 20 shows the Taqman results for an oncology panel (Phase II) of human tissues. 50226 expression was upregulated by in 6/6 colon tumor or colon metastases samples versus normal colon samples. 50226 expression was upregulated by in 5/6 lung tumor samples versus normal lung samples. 50226 expression was found in both breast and ovary tumors and normal breast and ovary samples.

TABLE 20


50226.2 Expression in Oncology Phase II Panel

Tissue Type	50226.2 Mean	β 2 Mean	∂∂ Ct	Expression

PIT 400 Breast N	29.77	20.4	9.38	1.50
PIT 372 Breast N	30.78	21.07	9.71	1.19
CHT 559 Breast N	35.76	22.82	12.94	0.00
MDA 304 Breast T: MD-IDC	30.93	19.61	11.32	0.39
CHT 2002 Breast T: IDC	29.42	20.3	9.13	1.79
MDA 236-Breast T:PD-IDC(ILC?)	32.06	20.57	11.48	0.35
CHT 562 Breast T: IDC	30.16	19.44	10.73	0.59
NDR 138 Breast T ILC (LG)	28.67	21.43	7.24	6.64
CHT 1841 Lymph node (Breast met)	33.12	22.2	10.91	0.52
PIT 58 Lung (Breast met)	33.91	23.27	10.64	0.63
CHT 620 Ovary N	27.41	20.14	7.27	6.48
PIT 208 Ovary N	26.11	19.45	6.66	9.89
CLN 012 Ovary T	29.97	22.59	7.38	5.98
CLN 07 Ovary T	29.81	19.73	10.07	0.93
CLN 17 Ovary T	27.32	20.9	6.42	11.64
MDA 25 Ovary T	30.21	22.52	7.7	4.83
MDA 216 Ovary T	29.27	19.29	9.98	0.99
PIT 298 Lung N	29.74	19.52	10.22	0.84
MDA 185 Lung N	32.22	20.52	11.71	0.30
CLN 930 Lung N	31.83	21.5	10.32	0.78
MPI 215 Lung T-SmC	27.18	19.74	7.45	5.74
MDA 259 Lung T-PDNSCCL	27.36	20.67	6.68	9.72
CHT 832 Lung T-PDNSCCL	29.51	19.36	10.15	0.88
MDA 262 Lung T-SCC	28.93	23.23	5.71	19.17
CHT 793 Lung T-ACA	26.51	19.22	7.29	6.37
CHT 331 Lung T-ACA	28.55	22.22	6.33	12.43
CHT 405 Colon N	28.07	17.4	10.66	0.62
CHT 523 Colon N	29.19	19.36	9.82	1.10
CHT 371 Colon N	26.55	16.68	9.86	1.08
CHT 382 Colon T: MD	29.84	21.75	8.1	3.64
CHT 528 Colon T: MD	25.31	18.47	6.84	8.73
CLN 609 Colon T	27.82	19.79	8.04	3.81
NDR 210 Colon T: MD-PD	30.75	24.11	6.64	9.99
CHT 340 Colon-Liver Met	28.59	21.66	6.92	8.23
NDR 100 Colon-Liver Met	25.22	18.55	6.67	9.82
PIT 260 Liver N (female)	28.43	19.15	9.29	1.60
CHT 1653 Cervix Squamous CC	27.37	21.91	5.46	22.72
CHT 569 Cervix Squamous CC	31.57	19.77	11.8	0.28
A24 HMVEC-Arr	27.13	19.58	7.54	5.35
C48 HMVEC-Prol	26.39	19.91	6.48	11.20
Pooled Hemangiomas	30.27	20.04	10.23	0.84
HCT116N22 Normoxic	25.3	22.16	3.14	113.44
HCT116H22 Hypoxic	26.24	22.8	3.44	91.82

The Taqman results in the following Table 21 show highest expression of 58764 orthologs in PBL HIV-1 d1.

TABLE 21


58764 (agpat) Expression

	58764	18S	relative exp.

PBL mock d1	26.6	12.9	0.50
PBL HIV-1 d1	24.4	11.3	0.75
PBL mock d3	26.3	11.9	0.31
PBL HIV-1 d3	25.7	11.7	0.40
PBL mock d5	28.1	12.6	0.14
PBL HIV-1 d5	27.3	12.1	0.17
M/M mock d7	29.0	12.4	0.06
M/M MOI 0.1 d7	29.3	12.1	0.04
M/M mock d15	29.2	12.2	0.05
M/M MOI 0.1 d15	30.2	14.1	0.10
M/M mock d26	31.1	14.7	0.08
M/M MOI 0.1 d26	30.6	12.4	0.02
Chronic-TRD-Pre	29.1	11.9	0.04
Chronic-TRD-Post	28.4	11.5	0.06
Chronic-MGB-Pre	29.0	12.1	0.05
Chronic-MGB-Post	28.9	11.6	0.04
Acute-EJT-Pre	28.3	11.7	0.07
Acute-EJT-Post	29.3	11.7	0.03
Acute-KEK-Pre	30.4	11.8	0.02
Acute-KEK-Post	28.9	11.4	0.03
Acute-RKY-Pre	31.2	12.1	0.01
Acute-RKY-Post	32.5	12.0	0.00
d2 uninfect.	29.4	13.7	0.13
d2 SIV239	29.6	12.9	0.06
d2 SIV316	28.0	12.1	0.10
d3 uninfect.	26.9	11.8	0.18
d3 SIV239	28.5	12.8	0.13
d3 SIV316	28.7	13.5	0.17
II AB	28.2	14.0	0.36
II M-CSF	30.2	14.7	0.14
IV AB	28.4	12.3	0.09
IV M-CSF	29.8	14.3	0.14

As seen by these results, 26199, 33530, 33949, or 50226 molecules have been found to be underexpressed or overexpressed in tumor cells, where the molecules may be inappropriately propagating either cell proliferation or cell survival signals. As such, activators or inhibitors of the 26199, 33530, 33949, or 50226 molecules are useful for the treatment of cancer; preferably ovarian, breast, colon, lung, liver or brain cancer, and useful as a diagnostic.

In situ Hybridization of 26199 and 33949

Specific in situ localization of gene 26199 was observed in 3/3 breast tumors and 1/1 ovarian tumor. No normal tissues for breast or ovary showed staining. 3/11 angiogenic tissues were also stained for 26199, including a Wilm's tumor and a neuroblastoma. Labeling was confined to epithelial cells with no evidence of expression by stromal blood vessels.

For 33949, 1/2 normal breast tissues, 4/5 breast tumors, 0/3 normal colon tissues, 3/4 primary colon tumors, 1/2 colon metastasis, 0/2 normal lung tissues, 2/4 lung tumors, 0/1 normal ovary tissue and 2/2 ovary tumors showed positive staining.

Human 62113

The present invention is based, in part, on the discovery of a novel acyl-CoA dehydrogenase family member, referred to herein as “62113”.

The human 62113 sequence (SEQ ID NO:53), which is approximately 3030 nucleotides long including untranslated regions, contains a predicted methionine-initiated coding sequence of about 2340 nucleotides, not including the termination codon (nucleotides 238-2577 of SEQ ID NO:53, 1-2340 of SEQ ID NO:55). The coding sequence encodes a 780 amino acid protein (SEQ ID NO:54).

Human 62113 contains the following regions or other structural features (for general information regarding PFAM identifiers, PS prefix and PF prefix domain identification numbers, refer to Sonnhammer et al. (1997) Protein 28:405-420): an acyl-CoA dehydrogenase N-terminal domain (PFAM Accession Number PF02771) located at about amino acid residues 483 to 503 of SEQ ID NO:54; an acyl-CoA dehydrogenase middle domains (PFAM Accession Number PF02770) located at about amino acid residues 505 to 585 of SEQ ID NO:54; an acyl-CoA dehydrogenase C-terminal domain (PFAM Accession Number PF00441) located at about amino acid residues 618 to 741 of SEQ ID NO:54; an acyl-CoA dehydrogenase signature 1-like sequence (Prosite PS00072) located at about amino acid residues 505 to 518 of SEQ ID NO:54; an acyl-CoA dehydrogenase signature 2-like sequence (Prosite PS00073) located at about amino acid residues 622 to 642 of SEQ ID NO:54; three N-glycosylation sites (Prosite PS00001) located at about amino acids 331 to 334, 499 to 502, and 558 to 561of SEQ ID NO:54; two glycosaminoglycan attachment sites (Prosite PS00002) located at about amino acids 34 to 37, and 540 to 543 of SEQ ID NO:54; one cAMP/cGMP-dependent protein kinase phosphorylation site (Prosite PS00004) located at about amino acids 774 to 777 of SEQ ID NO:54; six protein kinase C phosphorylation sites (Prosite PS00005) located at about amino acids 355 to 357, 376 to 378, 456 o 458, 655 to 657, 688 to 690, and 777 to 779 of SEQ ID NO:54; seven casein kinase II phosphorylation sites (Prosite PS00006) located at about amino acids 138 to 141, 275 to 278, 284 to 287, 333 to 336, 445 to 448, 507 to 510, and 517 to 520 of SEQ ID NO:54; nine N-myristoylation sites (Prosite PS00008) located at about amino acids 136 to 141, 170 to 175, 318 to 323, 330 to 335, 351 to 356, 498 to 503, 543 to 548, 699 to 704, 731 to 736 of SEQ ID NO:54; one amidation site (Prosite PS00009) located at about amino acids 533 to 536 of SEQ ID NO:54; one ATP/GTP-binding site motif A (P-loop; Prosite PS00017) located at about amino acids 47 to 54 of SEQ ID NO:54; one tyrosine protein kinase specific active-site signature sequence (Prosite PS00109) located at about amino acids 218 to 230 of SEQ ID NO:54; one microodies C-terminal targeting signal sequence (Prosite PS00342) located at about amino acids 778 to 781 of SEQ ID NO:54; and two eukaryotic thiol (cysteine) proteases histidien active site signature sequence (Prosite PS 00639) located at about amino acids 633 to 643 and 691 to 701 of SEQ ID NO:54.

A hydropathy plot of human 62113 was peformed. Polypeptides of the invention include fragments which include: all or part of a hydrophobic sequence; all or part of a hydrophilic sequence; and a sequence which includes a cysteine residue or a glycosylation site.

The 62113 protein contains a significant number of structural characteristics in common with members of the acyl-CoA dehydrogenase family. The acyl-CoA dehydrogenase family comprises a number of related enzymes that share high structural homology and a common catalytic mechanism which involves abstraction of an I-proton from the substrate (Thorpe and Kim (1995) FASEB J 9: 718-25). For example, acyl-CoA dehydrogenases catalyze the conversion of a fatty acyl thioester substrate to the corresponding I,θ-enoyl-CoA product. Thus, this family includes enzymes critical for the proper function of many physiological systems, including fatty acid oxidation, amino acid metabolism, and cellular proliferation and differentiation.

A 62113 polypeptide can include an “acyl-CoA dehydrogenase domain” or regions homologous with an “acyl-CoA dehydrogenase domain.”

As used herein, the term “acyl-CoA dehydrogenase domain” includes an amino acid sequence of about 50 to 500 amino acid residues in length, more preferably about 100 to 400 amino acid residues, or about 200 to 300 amino acids and has a bit score for the alignment of the sequence to the acyl-CoA dehydrogenase domain (HMM) of at least 4.9 or greater. The acyl-CoA dehydrogenase domain includes an amino acid sequence which has an all-alpha, four helical up-and-down bundle conformation at the C-terminal portion of the acyl-CoA dehydrogenase domain, e.g., an acyl-CoA dehydrogenase C-terminal domain (Pfam Accession Number PF 00441). The acyl-CoA dehydrogenase domain also includes an amino acid sequence which has a beta-barrel fold conformation and is found in the central domain of an acyl-CoA dehydrogenase, e.g., an acyl-CoA dehydrogenase middle domain (Pfam Accession Number PF02770). Preferably, the acyl-CoA middle domain includes an amino acid residue capable of providing a catalytic function to the active site, for example, an aspartate (D), at about amino acid 515 of SEQ ID NO:54. The acyl-CoA dehydrogenase domain also includes an amino acid sequence which has an all-alpha conformation and is found at the N-terminal portion of the acyl-CoA dehydrogenase domain, e.g., an acyl-CoA dehydrogenase N-terminal domain (Pfam Accession Number PF02771).

An alignment of the acyl-CoA dehydrogenase N-terminal domain (amino acids 483 to 503 of SEQ ID NO:54) of human 62113 with a consensus amino acid sequence derived from a hidden Markov model derived from Pfam (Pfam Accession Number PF02771; SEQ ID NO:56) has a bit score of 5.5 and E-value of 3.7.

An alignment of the acyl-CoA dehydrogenase middle domain (amino acids 505 to 585 of SEQ ID NO:54) of human 62113 with a consensus amino acid sequence derived from a hidden Markov model derived from Pfam (Pfam Accession Number PF02770; SEQ ID NO:57) has a bit score of 25.6 and E-value of 6.6e-06.

An alignment of the acyl-CoA dehydrogenase C-terminal domain (amino acids 618 to 741 of SEQ ID NO:54) of human 62113 with a consensus amino acid sequence derived from, a hidden Markov model derived from Pfam (Pfam Accession Number PF00441; SEQ ID NO:58) has a bit score of 24.6 and E-value of 1.5e-05.

A 62113 polypeptide can include a “acyl-CoA dehydrogenase domain”, e.g., an acyl-CoA dehydrogenase N-terminal domain, an acyl-CoA dehydrogenase middle domain, or an acyl-CoA dehydrogenase C-terminal domain, or regions homologous thereto.

As used herein, the term “acyl-CoA dehydrogenase N-terminal domain” includes an amino acid sequence of about 5 to 50 amino acid residues in length and having a bit score for the alignment of the sequence to the acyl-CoA dehydrogenase domain (HMM; Pfam Accession Number PF02771) of at least 5 and E-value of less than 4. Preferably, an acyl-CoA dehydrogenase N-terminal domain includes at least about 5 to 50 amino acids, more preferably about 10 to 40 amino acid residues, or about 15 to 25 amino acids and has a bit score for the alignment of the sequence to the acyl-CoA dehydrogenase N-terminal domain (HMM) of at least 3, preferably 4, more preferably 5, or greater and E-value of 6, preferably 5, more preferably 4 or less.

In a preferred embodiment, a 62113 polypeptide or protein has a “acyl-CoA dehydrogenase N-terminal domain” or a region which includes at least about 5 to 50, more preferably about 10 to 40, or 15 to 25 amino acid residues and has at least about 60%, 70%, 80%, 90%, 95%, 99%, or 100% homology with an “acyl-CoA dehydrogenase N-terminal domain,” e.g., the acyl-CoA dehydrogenase N-terminal domain of human 62113 (e.g., residues 483 to 503 of SEQ ID NO:54).

As used herein, the term “acyl-CoA dehydrogenase middle domain” includes an amino acid sequence of about 50 to 200 amino acid residues in length and having a bit score for the alignment of the sequence to the acyl-CoA dehydrogenase middle domain (HMM; Pfam Accession Number PF00441) of at least 20 and E-value of less than 1e-05. Preferably, an acyl-CoA dehydrogenase middle domain includes at least about 20 to 120 amino acids, more preferably about 50 to 100 amino acid residues, or about 75 to 90 amino acids and has a bit score for the alignment of the sequence to the acyl-CoA dehydrogenase middle domain (HMM) of at least 10, preferably 15, more preferably 20, or greater, and E-value of 1e-05, preferably 5e-05, more preferably 1e-06, or less.

The acyl-CoA dehydrogenase middle domain can include an acyl-CoA dehydrogenase signature 1 sequence pattern, or sequence patterns homologous thereto. A consensus sequence for this pattern is as follows: [GAC]-[LIVM]-[ST]-E-x(2)-[GSAN]-G-[ST]-D-x(2)-[GSA] (Prosite Accession No. PS00072; SEQ ID NO:59). The aspartate residue in the tenth position of the consensus sequence is a conserved residue in the active site of the enzyme and is important for its catalytic activity.

In the above conserved signature sequence, and other motifs or signature sequences described herein, the standard IUPAC one-letter code for the amino acids is used. Each element in the pattern is separated by a dash (-); square brackets ([]) indicate the particular residues that are accepted at that position; x indicates that any residue is accepted at that position; and numbers in parentheses (( )) indicate the number of residues represented by the accompanying amino acid.

A sequence pattern homologous to an acyl-CoA dehydrogenase signature 1 sequence pattern includes an acyl-CoA dehydrogenase signature 1-like sequence pattern which differs from the consensus sequence of Prosite Accession No. PS00072 by less than six, preferably less than five, more preferably less than four elements in the sequence. An acyl-CoA dehydrogenase signature 1-like sequence is located within the acyl-CoA dehydrogenase middle domain of human 62113 polypeptide, corresponding to amino acid residues at about 505 to 518 of SEQ ID NO:54. This acyl-CoA dehydrogenase signature 1-like sequence differs from an acyl-CoA dehydrogenase signature 1 sequence (e.g., Prosite Accession Number PS00072) in the fifth element, which consists of three amino acids, in the eighth element, which is a serine residue instead of glycine, and in the thirteenth element, which is an asparagine residue instead of glycine, serine, or alanine. The aspartate residue in the tenth position is a conserved residue and corresponds to the aspartate residue at about position 515 in SEQ ID NO:54.

In a preferred embodiment, a 62113 polypeptide or protein has an “acyl-CoA dehydrogenase middle domain” or a region which includes at least about 50 to 200, more preferably about 100 to 150, or about 115 to 135 amino acid residues and has at least about 60%, 70%, 80%, 90%, 95%, 99%, or 100% homology with an “acyl-CoA dehydrogenase middle domain,” e.g., the acyl-CoA dehydrogenase middle domain of human 62113 (e.g., residues 505 to 585 of SEQ ID NO:54).

In another preferred embodiment, a 62113 polypeptide or protein has an acyl-CoA dehydrogenase middle domain which includes an acyl-CoA dehydrogenase signature 1-like sequence pattern which differs from the consensus sequence (e.g., Prosite Accession No. PS00072) by less than six, preferably less than five, more preferably less than four elements in the sequence and has at least about 60%, 70%, 80%, 90%, 95%, 99%, or 100% homology with an acyl-CoA dehydrogenase signature 1-like sequence pattern, e.g., the acyl-CoA dehydrogenase signature 1-like sequence pattern of human 62113 (e.g., residues 505 to 518 of SEQ ID NO:54)

As used herein, the term “acyl-CoA dehydrogenase C-terminal domain” includes an amino acid sequence of about 50 to 200 amino acid residues in length and having a bit score for the alignment of the sequence to the acyl-CoA dehydrogenase domain (HMM) of at least 20 and E-value of less than 1e-04. Preferably, an acyl-CoA dehydrogenase C-terminal domain includes at least about 50 to 200 amino acids, more preferably about 80 to 150 amino acid residues, or about 110 to 140 amino acid residues and has a bit score for the alignment of the sequence to the acyl-CoA dehydrogenase domain (HMM) of at least 5, preferably 10, more preferably 15, or greater, and E-value of 1e-04, preferably 5e-04, more preferably 1e-05, or less.

The acyl-CoA dehydrogenase C-terminal domain can include an acyl-CoA dehydrogenase signature 2 sequence pattern, or sequence patterns homologus thereto. A consensus sequence for this pattern is as follows: [QDE]-x(2)-G-[GS]-x-G-[LIVMFY]-x(2)-[DEN]-x(4)-[KR]-x(3)-[DEN] (Prosite Accession No. PS00073; SEQ ID NO:60).

A sequence pattern homologous to an acyl-CoA dehydrogenase signature 2 sequence pattern includes an acyl-CoA dehydrogenase signature 2-like sequence pattern which differs from the consensus sequence of Prosite Accession No. PS00073 by less than six, preferably less than five, more preferably less than four elements in the sequence. An acyl-CoA dehydrogenase signature 2-like sequence is located within the acyl-CoA dehydrogenase C-terminal domain of human 62113 polypeptide, corresponding to amino acid residues at about 618 to 741 of SEQ ID NO:54. This acyl-CoA dehydrogenase signature 2-like sequence differs from an acyl-CoA dehydrogenase signature 2 sequence (e.g., Prosite Accession Number PS00073) in the second element, which consists of three amino acids, in the fifth element, which is an arginine residue instead of glycine or serine, in the eighth element, which is a proline residue instead of leucine, isoleucine, valine, methionine, phenylalanine, or tyrosine, and in the eleventh element, which is an isoleucine residue instead of aspartate, glutamate, or asparagine.

In a preferred embodiment, a 62113 polypeptide or protein has an “acyl-CoA dehydrogenase C-terminal domain” or a region which includes at least about 50 to 200, more preferably about 80 to 150, or about 110 to 140 amino acid residues and has at least about 60%, 70%, 80%, 90%, 95%, 99%, or 100% homology with an “acyl-CoA dehydrogenase C-terminal domain,” e.g., the acyl-CoA dehydrogenase C-terminal domain of human 62113 (e.g., residues 618 to 741 of SEQ ID NO:54).

In another preferred embodiment, a 62113 polypeptide or protein has an acyl-CoA dehydrogenase C-terminal domain which includes an acyl-CoA dehydrogenase signature 2-like sequence pattern which differs from-the consensus sequence (e.g., Prosite Accession No. PS00073) by less than seven, preferably less than six, more preferably less than five elements in the sequence and has at least about 60%, 70%, 80%, 90%, 95%, 99%, or 100% homology with an acyl-CoA dehydrogenase signature 2-like sequence pattern, e.g., the acyl-CoA dehydrogenase signature 1-like sequence pattern of human 62113 (e.g., residues 618 to 741 of SEQ ID NO:54).

To identify the presence of an “acyl-CoA dehydrogenase N-terminal domain”, an “acyl-CoA dehydrogenase middle domain”, or an “acyl-CoA dehydrogenase C-terminal domain” in a 62113 protein sequence, and make the determination that a polypeptide or protein of interest has a particular profile, the amino acid sequence of the protein can be searched against the Pfam database of HMMs (e.g., the Pfam database, version 6.6) using the default parameters. For example, the hmmsf program, which is available as part of the HMMER package of search programs, is a family specific default program for MILPAT0063 and a score of 15 is the default threshold score for determining a hit. Alternatively, the threshold score for determining a hit can be lowered (e.g., to 8 bits). A description of the Pfam database can be found in Sonhammer et al. (1997) Proteins 28:405-420 and a detailed description of HMMs can be found, for example, in Gribskov et al. (1990) Meth. Enzymol. 183:146-159; Gribskov et al. (1987) Proc. Natl. Acad. Sci. USA 84:4355-4358; Krogh et al. (1994) J. Mol. Biol. 235:1501-1531; and Stultz et al. (1993) Protein Sci. 2:305-314, the contents of which are incorporated herein by reference. A search was performed against the HMM database resulting in the identification of the “acyl-CoA dehydrogenase N-terminal domain” described above in the amino acid sequence of human 62113 at about residues 483 to 503 of SEQ ID NO:54, the “acyl-CoA dehydrogenase middle domain” described above in the amino acid sequence of human 62113 at about residues 505 to 585 of SEQ ID NO:54; and the “acyl-CoA dehydrogenase C-terminal domain” described above in the amino acid sequence of human 62113 at about residues 618 to 741 of SEQ ID NO:54.

A 62113 family member can include at least one acyl-CoA dehydrogenase N-terminal domain; at least one acyl-CoA dehydrogenase middle domain; and at least one acyl-CoA dehydrogenase C-terminal domain. A 62113 family member can include an acyl-CoA dehydrogenase middle domain with an acyl-CoA dehydrogenase signature 1-like sequence. A 62113 family member can also include an acyl-CoA dehydrogenase C-terminal domain with an acyl-CoA dehydrogenase signature 2-like sequence. Furthermore, a 62113 family member can include at least one, two, preferably three N-glycosylation site (Prosite PS00001); at least one, preferably two glycosaminoglycan attachment sites (Prosite PS00002); at least one cAMP/cGMP protein kinase phosphorylation sites (Prosite PS00004); at least one, two, three, four, five, preferably six protein kinase C phosphorylation sites (Prosite PS00005); at least one, two, three, four, five, six, preferably seven [as appropriate] casein kinase II phosphorylation sites (Prosite PS00006); at least one, two, three, four, five, six, seven, eight, preferably nine N-myristoylation sites (Prosite PS00008); at least one amidation site (Prosite PS00009); at least one ATP/GTP binding site motif A (P-loop; Prosite PS00017); at least one tyrosine protein kinase specific active site signature sequence(Prosite PS00109); at least one microbodies C-terminal targeting signal (Prosite PS00342); and at least one, preferably two eukaryotic thiol (cysteine) protease histidine active site (Prosite PS00639).

Based on the above-described sequence similarities, the 62113 molecules of the present invention are predicted to have similar biological activities as acyl-CoA dehydrogenase family members. For example, the 62113 protein is predicted to have one or more of the following activities: (1) the ability to catalyze the transfer of hydrogen and electrons from one compound to another; (2) the ability to catalyze the I,θ-dehydrogenation of fatty acyl-CoA derivatives; (3) the ability to catalyze the dehydrogenation of branched short-chain acyl-CoAs in the metabolism of the branched-chain amino acids; (4) the ability to modulate the oxidation of fatty acids; (5) the ability to modulate the metabolism of amino acids; (6) the ability to modulate a cardiovascular activity; (7) the ability to modulate a renal activity; or (8) the ability to modulate a hepatic activity. As a result, the 62113 protein may have a critical function in one or more of the following physiological processes: (1) fatty acid metabolism; (2) amino acid metabolism; (3) modulation (stimulation or inhibition) of cell proliferation and differentiation; (4) modulation of tumorigenesis and tumor invasion; (5) cardiovascular activity; (6) renal activity; or (7) hepatic activity.

As the 62113 polypeptides of the invention can modulate 62113-mediated activities, they can be useful for developing novel diagnostic and therapeutic agents for acyl-CoA dehydrogenase-associated or other 62113-associated disorders, as described below.

As used herein, a “62113 activity”, “biological activity of 62113” or “functional activity of 62113”, refers to an activity exerted by a 62113 protein, polypeptide or nucleic acid molecule on e.g., a 62113-responsive cell or on a 62113 substrate, e.g., a protein substrate, as determined in vivo or in vitro. In one embodiment, a 62113 activity is a direct activity, such as an association with a 62113 target molecule. A “target molecule” or “binding partner” is a molecule with which a 62113 protein binds or interacts in nature. In an exemplary embodiment, 62113 is an enzyme that metabolizes fatty acyl-CoA substrates.

A 62113 activity can also be an indirect activity, e.g., a cellular signaling activity mediated by interaction of the 62113 protein with a 62113 receptor.

The 62113 molecules of the invention can modulate the activities of cells in tissues where they are expressed. For example, 62113 mRNA is expressed in tumors and metastases of the lung, colon, prostate, breast, and ovaries; heart, kidney, liver, and prostate. Accordingly, the 62113 molecules of the invention can act as therapeutic or diagnostic agents for tumors and metastases of the lung, colon, prostate, breast, and ovaries; and cardiovascular, renal, hepatic, and prostate disorders.

Examples of cellular proliferative and/or differentiative disorders include cancer, e.g., carcinoma, sarcoma, metastatic disorders or hematopoietic neoplastic disorders, e.g., leukemias. A metastatic tumor can arise from a multitude of primary tumor types, including but not limited to those of prostate, colon, lung, breast, ovarian, and liver origin.

The 62113 molecules of the invention can be used to monitor, treat and/or diagnose a variety of proliferative disorders. Such disorders include hematopoietic neoplastic disorders.

Gene Expression Analysis of 62113

Human 62113 expression was measured by TaqMan® quantitative PCR (Perkin Elmer Applied Biosystems) in cDNA prepared from a variety of normal and diseased (e.g., cancerous) human tissues or cell lines.

The results indicate significant 62113 expression in normal liver and in liver fibrosis samples; normal breast and breast tumor; normal ovary and ovarian tumor; normal prostate and prostate tumor; upregulated expression in lung tumor (e.g., adenocarcinoma) when compared to normal lung; upregulated expression in colon tumor (e.g., adenocarcinoma) when compared to normal colon; upregulated expression in diseased heart when compared to normal heart; and upregulated expression in diseased kidney (including hypertensive kidney) when compared to normal kidney.

Additional TaqMan panels showed the following patterns of 62113 expression: high levels of 62113 expression in HUVEC (human umbilical vein endothelial cells), normal adrenal gland, normal brain cortex and brain hypothalamus, dorsal root ganglia, and diseased aoartic tissue; and moderate levels of 62113 expression in pancreas, normal spinal cord and hemangiomas.

Human 32144

The present invention is based, in part, on the discovery of a novel fatty acid amide hydrolase family member, referred to herein as “32144”.

The human 32144 sequence (SEQ ID NO:61), which is approximately 2004 nucleotides long including untranslated regions, contains a predicted methionine-initiated coding sequence of about 1596 nucleotides, (nucleotides 119-1714 of SEQ ID NO:61; 1-1596 of SEQ ID NO:63) not including the termination codon. The coding sequence encodes a 532 amino acid protein (SEQ ID NO:62).

Portions of the fatty acid amide hydrolase domain of human 32144 align with consensus amino acid sequences derived from a hidden Markov model (HMM) from PFAM. The two distinct and non-overlapping consensus amino acid sequences correspond to portions of the PFAM amidase domain, PF01425. The scores for the two individual alignments were: 219.6 (E-value=4.8e-62) and 38.4 (E-value=6.6e-10), and the combined score for the two alignments was: 258.0 (E-value=1.3e-73). In the first alignment, the consensus amino acid sequence (residues 1-218 of the domain or SEQ ID NO:64) of an N-terminal portion of the amidase domain aligns with amino acids 69 to 289 of SEQ ID NO:62. In the second alignment, the consensus amino acid sequence (residues 395-521 of the domain or SEQ ID NO:65) of a C-terminal portion of the amidase domain aligns with amino acids 419 to 513 of SEQ ID NO:62.

Human 32144 contains the following regions or other structural features: an amidase domain (PFAM- Accession Number PF01425) located at about amino acid residues 69 to 289 and 419 to 513 of SEQ ID NO:62; an amidase signature motif (PS00571) located at about amino acid residues 204 to 235 of SEQ ID NO:62; a transmembrane domain located at about amino acid residues 11 to 33 of SEQ ID NO:62; eight predicted Protein Kinase C phosphorylation sites (PS00005) located at about amino acid residues 6 to 8, and 40 to 42, 129 to 131, 186 to 188, 230 to 232, 329 to 331, 365 to 367, and 434 to 436 of SEQ ID NO:62; three predicted Casein Kinase II phosphorylation sites (PS00006) located at about amino acid residues 129 to 132, 207 to 210, and 320 to 323 of SEQ ID NO:62; eleven predicted N-myristoylation sites (PS00008) located at about amino acid residues 53 to 58, 125 to 130, 138 to 143, 172 to 177, 204 to 209, 211 to 216, 224 to 229, 248 to 253, 475 to 480, 481 to 486, and 495 to 500 of SEQ ID NO:62; two predicted N-glycosylation sites (PS00001) at about amino acids 141 to 144 and 175 to 178 of SEQ ID NO:62; and one predicted microbodies C-terminal targeting signal (PS00342) at about amino acid 530 to 532 of SEQ ID NO:62.

A hydropathy plot of human 32144 was performed. Polypeptides of the invention include fragments which include: all or part of a hydrophobic sequence, e.g., the sequence from about amino acid 157 to 182, from about 388 to 414, and from about 471 to 491 of SEQ ID NO:62; all or part of a hydrophilic sequence, e.g., the sequence of from about amino acid 104 to 120, from about 183 to 201, and from about 415 to 438 of SEQ ID NO:62.

The 32144 protein contains a significant number of structural characteristics in common with members of the amidase family. An amidase family of proteins, also referred to as fatty acid amidase hydrolases (FAAH), is characterized by the ability to hydrolyze fatty acid amides, e.g., neuromodulatory fatty acid amides, such as oleamide, anandamide and myristic amide. Representative amidases include fatty acid amide hydrolases (FAAH) from human and mouse (Giang, D. K. et al. (1997) Proc. Natl. Acad. Sci. 94: 2238-2242). Typically, amidases possess substrate specificity based on chain length and degree of saturation of fatty acid amides. Fatty acid amides, e.g., oleamide and ananadmide, are known to have sleep-inducing and analgesic properties, as well as the ability to regulate cellular proliferation. This family of proteins typically contains a highly conserved region rich in glycine, serine and alanine residues. Fatty acid amide hydrolases have been described in Ueda et al. (2000), supra, the contents of which are incorporated herein by reference.

A 32144 polypeptide can include at least one “amidase domain” or “fatty acid amid hydrolase domain”, which contains one and preferably two “amidase subdomains” or regions homologous with an “amidase domain”.

As used herein, the term “amidase subdomain” or “first amidase subdomain” includes an amino acid sequence of about 100 to 500 amino acid residues in length and having a bit score for the alignment of the sequence to the amidase domain (HMM) of at least 100. Preferably, an amidase domain includes at least about 150 to 450 amino acids, more preferably about 200 to 300 amino acid residues, or about 220 amino acids and has a bit score for the alignment of the sequence to the amidase domain (HMM) of at least 150, preferably 200 or greater. The amidase domain (HMM) has been assigned the PFAM Accession Number PF01425. The first amidase domain (amino acids 69 to 289 of SEQ ID NO:62) of human 32144 aligns with a consensus amino acid sequence derived from a hidden Markov model.

The term “amidase subdomain” or “second amidase subdomain” includes an amino acid sequence of about 40 to 300 in length and having a bit score for the alignment of the sequence to the amidase domain (HMM) of at least 10. Preferably, an amidase domain includes at least about 60 to 200 amino acids, more preferably about 80 to 100 amino acid residues, or about 94 amino acids and has a bit score for the alignment of the sequence to the amidase domain (HMM) of at least 20, preferably 30 or greater. The amidase domain (HMM) has been assigned the PFAM Accession Number PF01425. The second amidase subdomain (amino acid residues 419 to 513 of SEQ ID NO:62) of human 32144 aligns with a consensus amino acid sequence derived from a hidden Markov model.

In a preferred embodiment, a 32144 polypeptide or protein has at least one “amidase subdomain” or a region that includes at least the size ranges described above and has at least about 60%, 70% 80% 90% 95%, 99%, or 100% homology with an “amidase domain,” e.g.; the amidase subdomain of human 32144 (e.g., residues 69 to 289 or 419 to 513 of SEQ ID NO:62).

To identify the presence of an “amidase” or “fatty acid amide hydrolase” domain in a 32144 protein sequence, and make the determination that a polypeptide or protein of interest has a particular profile, the amino acid sequence of the protein can be searched against the PFAM database of HMMs (e.g., the Pfam database, release 2.1) using the default parameters. For example, the hmmsf program, which is available as part of the HMMER package of search programs, is a family specific default program for MILPAT0063 and a score of 15 is the default threshold score for determining a hit. Alternatively, the threshold score for determining a hit can be lowered (e.g., to 8 bits). A description of the PFAM database can be found in Sonhammer et al. (1997) Proteins 28(3):405-420 and a detailed description of HMMs can be found, for example, in Gribskov et al.(1990) Meth. Enzymol. 183:146-159; Gribskov et al.(1987) Proc. Natl. Acad. Sci. USA 84:4355-4358; Krogh et al.(1994) J. Mol. Biol. 235:1501-1531; and Stultz et al.(1993) Protein Sci. 2:305-314, the contents of which are incorporated herein by reference. A search was performed against the HMM database resulting in the identification of a “amidase” domain in the amino acid sequence of human 32144, which includes two amidase subdomains located at about amino acid residues 69 to 289 and 419 to 513 of SEQ ID NO:62.

In one embodiment, a 32144 protein includes at least one amidase signature motif. As used herein, an “amidase signature motif” includes a sequence of at least nineteen amino acid residues defined by the sequence: G-[G/A]-S-[G/S]-[G/S]-G-X-[G/S/A]-[G/S/A/V/Y]-X-[G/A]-X-[D/E]-X-[G/A]-X-S-[L/I/V/M]-R—X—P-[G/S/A/C] (SEQ ID NO:66). An amidase signature motif, as defined, can be involved in the enzymatic hydrolysis of a fatty acid amide. More preferably, an amidase signature motif includes 25, 29, or even more preferably 32 amino acid residues. Amidase signature motifs have been described in, e.g., Mayaux et al. (1990), J Bacteriology 172:6764-73, the contents of which are incorporated herein by reference. Human 32144 contains a sequence (about amino acid residues 204-235 of SEQ ID NO:62) which matches the sequence of an amide signature motif at 18/19 of the conserved positions. The single discrepancy occurs at position 9 ([G/S/A/V/Y]) of the amidase signature sequece, where there is a conservative cystein substitution (located at about amino acid residue 212 of SEQ ID NO:62) observed in human 32144.

In a preferred embodiment, a 32144 polypeptide or protein has at least one amidase signature motif, or a region which includes at least 19, 25, 29, or even 32 amino acid residues and has at least 70%, 80%, 90%, or 100%-homology with an “amidase signature motif” or the variant amidase signature motif observed in human 32225, e.g., about amino acid residues 204 to 235 of SEQ ID NO:62.

A 32144 molecule can further include a transmembrane region. As used herein, the term “transmembrane domain” includes an amino acid sequence of at least about 14 amino acid residues in length that spans a phospholipid membrane. More preferably, a transmembrane domain includes at least about 14, 16, 18, 20, 22, or 24 amino acid residues and spans a phospholipid membrane. Transmembrane domains are rich in hydrophobic residues, and typically have an I-helical structure. In a preferred embodiment, at least 50%, 60%, 70%, 80%, 90%, 95% or more of the amino acids of a transmembrane domain are hydrophobic, e.g., leucines, valines, alanines, phenylalanines, methionines, isoleucines, tyrosines, or tryptophans. Transmembrane domains are described in, for example, Zagotta W. N. et al., (1996) Annual Rev. Neuronsci. 19:235-63.

In a preferred embodiment, a 32144 polypeptide or protein has at least one transmembrane domain or a region which includes at least 18, 19, or 20 amino acid residues and has at least about 60%, 70%, 80%, 90%, 95%, 99%, or 100% homology with a “transmembrane domain,” e.g., at least one transmembrane domain of human 32144 (e.g., from about amino acid residues 11 to33 of SEQ ID NO:62). In one embodiment, the transmembrane domain of a 32144 molecule is able to interact with transmembrane domains of other molecules, e.g. other 32144 molecules, such that the 32144 forms an oligomer, e.g., a homooligomer. The self-association of fatty acid amide hydrolases via N-termial transmembrane domains has been described in Ueda et al. (2000), supra.

A 32144 family member can include at least one, and preferably two amidase subdomains. Furthermore, a 32144 family member can include at least one amidase signature motif; at least one transmembrane domain; at least one, two, three, four, five, six, seven, and preferably eight predicted protein kinase C phosphorylation sites (PS00005); at least one, two, and preferably three predicted casein kinase II phosphorylation sites (PS00006); at least one, two, three, four, five, six, seven, eight, nine, ten, and preferably eleven predicted N-myristylation sites (PS00008); at least one, and preferably two predicted N-glycosylation sites (PS00001); and at least one predicted Microbodies C-terminal targeting signal (PS00342).

As the 32144 polypeptides of the invention may modulate 32144-mediated activities, they may be useful as of for developing novel diagnostic and therapeutic agents for 32144-mediated or related disorders, as described below.

As used herein, a “32144 activity”, “biological activity of 32144” or “functional activity of 32144”, refers to an activity exerted by a 32144 protein, polypeptide or nucleic acid molecule For example, a 32144 activity can be an activity exerted by 32144 in a physiological milieu on, e.g., a 32144-responsive cell or on a 32144 substrate, e.g., a protein substrate. A 32144 activity can be determined in vivo or in vitro. In one embodiment, a 32144 activity is a direct activity, such as an association with a 32144 target molecule. A “target molecule” or “binding partner” is a molecule with which a 32144 protein binds or interacts in nature. In an exemplary embodiment, 32144 is an enzyme that hydrolyses fatty acid amides, e.g., anandamide or ethanolamides of oleic (e.g., oleamide), linoleic, or palmitic acids.

A 32144 activity can also be an indirect activity, e.g., a cellular signaling activity mediated by interaction of the 32144 protein with a 32144 receptor. The features of the 32144 molecules of the present invention can provide similar biological activities as fatty acid amide hydrolase family members. For example, the 32144 proteins of the present invention can have one or more of the following activities: (1) bind and catabolize fatty acid amides; (2) regulate neuronal signaling; (3) regulate ion channel function, e.g., 5-HT ₃ion channel function; (4) regulate cannabinoid receptor signaling; (5) regulate seratonin signaling, e.g., 5-HT₂response to seratonin; (6) regulate gap junction activity; (7) regulate pain reception; (8) regulate development; (9) regulate cellular proliferation and/or migration; (10) regulate focal adhesion kinase activity; or (11) regulate the induction of sleep.

Thus, the 32144 molecules can act as novel diagnostic targets and therapeutic agents for controlling cellular proliferation and/or differentiation disorders, disorders of the brain, CNS, or peripheral nervous system, metabolic and pain disorders, or sleep disorders, e.g., narcolepsy.

Examples of cellular proliferation and/or differentiation disorders include cancer, e.g., carcinoma, sarcoma, metastatic disorders or hematopoietic neoplastic disorders, e.g., leukemias. A metastatic tumor can arise from a multitude of primary tumor types, including but not limited to those of prostate, colon, lung, breast and liver origin.

Tissue Distribution of 32144 mRNA

Endogenous human 32144 gene expression was determined using the Perkin-Elmer/ABI 7700 Sequence Detection System which employs TaqMan technology. Tissues tested include the human tissues and several cell lines shown in Tables 22-25. 32144 mRNA was detected in a number of tissues, including the kidney, pancreas, brain, and liver (Table 22). Importantly, 32144 expression was upregulated in most of the lung, colon, breast, and ovarian tumors tested (Tables 22-24). 32144 mRNA was also detected in several tumor cell lines, whether grown in vivo (Table 25) or in vitro (Table 26), and growth of breast tumor cell lines on agar correlated with increased expression of 32144 mRNA as compared to growth on plastic (Table 26).

The incidence of tumor-associated expression of 32144 mRNA in lung, ovary, breast, and colon tissues was further evaluated by in situ hybridization (Table 27). Notable tumor-associated expression of 32144 is seen in all of the different tumor types tested. This data, like the Taqman data, suggests a role for 32144 in tumor development. In addition, expression of 32144 mRNA in invasive indolent breast carcinomas vs. metastatic breast carcinomas was evaluated by hybridizing tumor cell RNA to microarray chips that were capable of detecting 32144 nucleic acids (Table 28). All of the tumors tested expressed 32144 mRNA, while 2/5 metastatic tumors and 0/3 invasive indolent tumors displayed a relative increase in 32144 expression. This data, along with the colon tumor in-situ hybridization data reveals a positive correlation between 32144 expression and tumor metastasis, at least for breast and colon tumors.

TABLE 22


			Relative
Tissue Type	32144 Mean	β 2 Mean	Expression

Artery normal	34.72	22.38	0.19
Aorta diseased	33.33	23.00	0.78
Vein normal	35.26	20.66	0.00
Coronary SMC	35.06	21.18	0.00
HUVEC	31.24	21.50	1.17
Hemangioma	31.99	20.05	0.25
Heart normal	33.42	20.90	0.17
Heart CHF	31.20	20.26	0.51
Kidney	27.57	20.53	7.60
Skeletal Muscle	40.00	28.39	0.00
Adipose normal	37.55	29.80	0.00
Pancreas	28.33	22.43	16.69
primary osteoblasts	35.35	21.07	0.00
Osteoclasts (diff)	34.77	17.92	0.01
Skin normal	31.45	22.34	1.81
Spinal cord normal	32.28	21.36	0.52
Brain Cortex normal	29.65	22.56	7.37
Brain Hypothalamus normal	30.93	22.57	3.02
Nerve	34.01	22.24	0.29
DRG (Dorsal Root Ganglion)	33.84	22.35	0.35
Breast normal	30.32	21.55	2.28
Breast tumor	28.50	20.88	5.05
Ovary normal	29.64	20.07	1.32
Ovary Tumor	29.09	20.02	1.85
Salivary glands	28.74	19.91	2.19
Colon normal	28.41	18.40	0.97
Colon Tumor	28.68	21.98	9.62
Lung normal	28.62	18.32	0.79
Lung tumor	25.91	20.45	22.72
Lung COPD	28.17	18.70	1.41
Colon IBD	27.64	18.00	1.26
Liver normal	27.31	20.34	7.98
Liver fibrosis	27.41	20.89	10.82
Spleen normal	30.79	19.91	0.53
Tonsil normal	25.54	17.77	4.60
Lymph node normal	28.48	19.77	2.40
Small intestine normal	30.61	20.50	0.91
Macrophages	31.80	17.40	0.05
Synovium	31.52	19.78	0.29
BM-MNC	33.83	19.09	0.04
Activated PBMC	28.09	18.29	1.12
Neutrophils	36.31	19.26	0.00
Megakaryocytes	32.13	19.11	0.12
Erythroid	33.20	22.05	0.44
positive control	28.92	20.59	3.12

As shown in the “Relative Expression” column of Table 22, 32144 mRNA is expressed in the pancreas, kidney, liver, cerebral cortex, hypothalamus, tonsils, lymph nodes, breast, salivary gland, skin, and ovary. Weak expression is observed in the heart and blood vessels, dorsal root gaglia, colon, lung, spleen, small intestine, and blood cells. In addition, 32144 expression is highly upregulated in lung, colon, and breast tumors, and slightly upregulated in ovarian tumors. Abbreviations used in Table 22: SMC, smooth muscle cell; HUVEC, human umbilical vein endothelial cells; CHF, congestive heart failure; diff, differentiated; COPD, chronic obstructive pulmonary disease; IBD, inflammatory bowel disease; BM-MNC, bone marrow mononuclear cell; PBMC, pre-bone marrow cell.

TABLE 23


	32144.1		Relative
Tissue Type	Mean	β 2 Mean	Expression

PIT 400 Breast N	31.85	20.18	0.31
PIT 372 Breast N	32.09	20.92	0.43
PIT 271 Breast N	35.4	25.48	0.00
MDA 106 Breast T	31.93	21.11	0.55
MDA 234 Breast T	29.27	18.77	0.69
NDR 57 Breast T	30.98	19.75	0.41
MDA 304 Breast T	29.8	19.34	0.71
NDR 58 Breast T	26.43	17.88	2.66
NDR 132 Breast T	30.15	21.54	2.57
NDR 07 Breast T	30	19.65	0.76
NDR 12 Breast T	28.84	21.69	7.02
PIT 208 Ovary N	32.35	19.52	0.14
CHT 620 Ovary N	34.23	20.1	0.06
CHT 619 Ovary N	34.7	20.6	0.06
CLN 03 Ovary T	29.57	20.08	1.39
CLN 17 Ovary T	27.84	20.35	5.58
CLN 07 Ovary T	30.43	19.66	0.57
CLN 08 Ovary T	29.87	18.9	0.50
MDA 216 Ovary T	33.05	21.04	0.24
CLN 012 Ovary T	31.83	22.16	1.22
MDA 25 Ovary T	29.89	22.62	6.50
MDA 183 Lung N	30.52	18.43	0.23
CLN 930 Lung N	32.03	19.36	0.15
MDA 185 Lung N	34.11	19.88	0.05
CHT 816 Lung N	30.36	17.2	0.11
MPI 215 Lung T-SmC	31.25	18.91	0.19
MDA 259 Lung I-PDNSCCL	27.37	19.87	5.52
CHT 832 Lung T-PDNSCCL	28.59	19.36	1.68
MDA 253 Lung T-PDNSCCL	29.66	19	0.62
CHT 911 Lung T-SCC	28.73	19.3	1.45
CHT 793 Lung T-ACA (?)	29.3	19.2	0.91
MDA 262 Lung T-SCC	31.18	23.35	4.38
CHT 211 Lung T-AC	28.01	19.86	3.53
NHBE	30.45	21.66	2.27
MDA 127 N Ovarian	34.08	16.97	0.01
Epithelial Cells
MDA 224 N Ovarian	36.37	16.62	0.00
Epithelial Cells
MDA 124 Ovarian Ascites	28.13	15.52	0.16
MDA 126 Ovarian Ascites	26.59	17.4	1.71

As shown in the “Relative Expression” column of Table 23, 32144 mRNA expression is slightly upregulated in 4/8 of the breast tumor samples tested, as compared to normal breast tissue, and dramatically upregulated in 3/8 of the breast tumor samples. Likewise, 7/7 ovary tumor samples displayed an increase in 32144 expression relative to normal ovary tissue, while 2/7 contained dramatically upregulated levels of 32144 mRNA. Amongst lung tumor samples tested, 8/9 displayed an increase in 32144 expression relative to normal lung tissue, with 4/9 containing highly elevated levels of 32144 mRNA. Abbreviations used in Table 23: N, normal tissue; T, tumor; SmC, small cell carcinoma; PDNSCCL, poorly differentiated non-small cell carcinoma; SCC, squamous cell carcinoma; AC, adenocarcinoma; NHBE, lung cell line.

TABLE 24


			Relative
Tissue Type	32144.1 Mean	β 2 Mean	Expression

CHT 523 Colon N	31.64	19	0.16
NDR 104 Colon N	28.39	19.09	1.59
CHT 416 Colon N	30.23	19.48	0.58
CHT 452 Colon N	32	18.7	0.10
NDR 210 Colon T	32.7	24.07	2.51
CHT 398 Colon T	27.32	19.91	5.84
CHT 382 Colon T	26.62	18.97	4.96
CHT 944 Colon T	28.61	18.84	1.15
CHT 528 Colon T	26.5	19.07	5.78
CHT 1365 Colon T	27.56	19.2	3.05
CHT 372 Colon T	30.31	20.27	0.95
CLN 609 Colon T	28.55	20.02	2.70
CHT 01 Liver Met	28.29	18.11	0.87
NDR 100 Liver Met	26.37	18.59	4.53
CHT 340 Liver Met	30.07	20.79	1.60
NDR 217 Liver Met	29.04	19.2	1.10
PIT 260 Liver N	27.5	17.77	1.17
CHT 320 Liver N	30.67	23.56	7.21
C48 HMVEC-Prol	36.87	20.74	0.00
ONC 102 Hemangioma	33.59	20.15	0.09

As shown in the “Relative Expression” column of Table 24, 6/8 of the tested colon tumors had an elevated level of 32144 expression as compared to normal colon tissue, with 3/8 displaying a dramatic increase in 32144 mRNA expression. All liver metastases tested expressed 32144 mRNA. Abbreviations used in Table 24: N, normal tissue; T, tumor; Met, metastasis; HMVEC, human vascular endothelial cells; prol, proliferating.

TABLE 25


	32144.1	B-2	Relative
Cell Line	Mean	Mean	Expression

MCF-7 Breast T	25.21	18.88	12.43
ZR75 Breast T	26.34	20.29	15.09
T47D Breast T	25.98	18.00	3.96
MDA 231 Breast T	33.38	17.35	0.01
MDA 435 Breast T	30.20	15.89	0.05
SKBr3 Breast	28.63	18.99	1.25
DLD 1 ColonT (stageC)	24.36	19.44	33.03
SW620 ColonT (stageC)	25.12	18.26	8.58
HCT116	25.33	18.00	6.22
HT29	25.09	15.88	1.68
Colo 205	24.23	14.83	1.48
NCIH125	30.26	17.38	0.13
NCIH322	25.66	18.36	6.37
NCIH460	32.71	17.33	0.02
A549	32.39	18.62	0.07
NHBE	30.14	21.37	2.29
SKOV-3 ovary	28.29	17.24	0.47
OVCAR-3 ovary	26.55	20.31	13.23
293 baby kidney	27.04	20.28	9.23
293T baby kidney	32.24	21.60	0.63

Table 25 depicts the relative expression of 32144 mRNA in cell lines that have been xenographed into mice and allowed to form tumors. Several of the lines display high levels of 32144 expression when grown under such conditions. Most notable is one of the Stage C colon tumor lines, a couple of the breast tumor lines, one of the ovary carcinoma lines, and a baby kidney fibroblast line. Many of the other cell lines also express 32144 mRNA when xenographed into mice. Abbreviation used in Table 25: T, tumor; HCT116, HT29, and Colo 205, colon carcinoma cell lines; NCIH125, NCIH322, NCIH460, A549, and NHBE, lung carcinoma cell lines.

TABLE 26


	32144.1		Relative
Tissue Type	Mean	β 2 Mean	Expression

MCF10MS	31.31	19.84	0.35
MCF10A	37.33	19.75	0.00
MCF10AT.cl1	39.96	19.48	0.00
MCF10AT.cl3	38.22	18.86	0.00
MCF10AT1	31.68	19.94	0.29
MCF10AT3B	39.88	19.47	0.00
MCF10CA1a.cl1	34.26	17.09	0.01
MCF10AT3B Agar	32.53	25.9	10.10
MCF10CA1a.cl1 Agar	33.95	24.5	1.43
MCF10A.m25 Plastic	37.11	24.54	0.00
MCF10CA Agar	33.07	21.5	0.33
MCF10CA Plastic	33.27	21.56	0.30
MCF3B Agar	29.58	21.84	4.68
MCF3B Plastic	30.32	21.58	2.35
MCF10A EGF 0 hr	32.59	17.23	0.02
MCF10A EGF 0.5 hr	32.2	17.45	0.04
MCF10A EGF 1 hr	32.63	17.6	0.03
MCF10A EGF 2 hr	32.83	17.63	0.03
MCF10A EGF 4 hr	33.52	17.63	0.02
MCF10A EGF 8 hr	33.31	17.52	0.02
MCF10A IGF1A 0 hr	31.05	21.58	1.41
MCF10A IGF1A 0.5 hr	31.36	21.75	1.27
MCF10A IGF1A 1 hr	30.93	21.84	1.84
MCF10A IGF1A 3 hr	30.78	21.88	2.09
MCF10A IGF1A 24 hr	29.32	21.84	5.62
MCF10AT3B.cl5 Plastic	35.42	21.82	0.00
MCF10AT3B.cl6 Plastic	35.7	21.85	0.00
MCF10AT3B.cl3 Plastic	36.19	21.63	0.00
MCF10AT3B.cl1 Plastic	35.02	21.72	0.00
MCF10AT3B.cl4 Plastic	35.09	21.47	0.00
MCF10AT3B.cl2 Plastic	36.45	21.84	0.00
MCF10AT3B.cl5 Agar	31.91	24.06	4.33
MCF10AT3B.cl6 Agar	32.32	24.05	3.23
MCF-7	30.1	23.27	8.76
ZR-75	28.29	21.59	9.65
T47D	29.61	21.64	3.97
MDA-231	36.9	20.45	0.00
MDA-435	36.16	20.55	0.00
SkBr3	30.81	20.93	1.06
Hs578Bst	36.78	19.85	0.00
Hs578T	38.49	19.66	0.00

Table 26 depicts the relative expression of 32144 mRNA in breast carcinoma cell lines grown under various conditions. Growth of the cell lines on agar correlates with an increase in 32144 expression, as shown by the MCF10AT3B, MCF3B, MCF10AT3B clone 5 and MCT10AT3B clone 6 cell lines. MCF10A cells did not display a change in 32144 expression in response to epidermal growth factor (EGF), while they did respond to insulin growth factor 1A (IGF-1A) by gradually increasing 32144 mRNA expression over the course of 24 hours.

TABLE 27


Spectrum	Tissue	Expression

CHT 800	Lung—PD-NSC	+/−
CHT 813	Lung—MD-SCC	−/−
CHT 726	Lung—MD-SCC	+/−
CHT 331	Lung—MD-AC	−/−
MPI 216	Lung—Normal	−/−

(LUNG: 0/1 normals; 2/4 tumors)

MDA 28	Ovary—Malignant	−/−
MDA 300	Ovary—MD-AC	+/−
MDA 202	Ovary—Normal	−/−

(OVARY: 0/1 normals; 1/2 tumors)

NDR 7	Breast—IDC	+/−
NDR 12	Breast—IDC	+++/+
NDR 57	Breast—PD-Ductal AC	+/−
CLN 662	Breast—IDC/IDL	+/−
MDA 156	Breast—DCIS	++/+
CLN 156	Breast—PD-IDC	+/+
MDA 91	Breast—AC	+++/+
PIT 58	Breast—PD-AC(lung)	−/−
CHT 1841	Breast—Met AC(lymph)	+/−
PIT 116	Breast—Met AC(lung)	+/−
MDA 405	Breast—normal	−/−

(BREAST: 0/1 normals; 7/7 tumors; 2/3 metastasis)

CLN 609	Colon—Invasive	−/−
NDR 99	Colon—Invasive	+/−
NDR 100	Colon—AC(liver)	+/+
CHT 1	Colon—Met AC	++/+

	(Colon: 1/2 tumors; 2/2 metastasis)

Expression of 32144 mRNA was analyzed by in-situ hybridization in both normal and tumor tissue samples. Expression of 32144 mRNA was consistently observed in the tumors, suggesting a role for 32144 in tumor development. Furthermore, in colon tumor samples, expression of 32144 mRNA was more prevalent in metastatic tumors, indicating a possible link between 32144 expression and tumor metastasis in some tissues. Abbreviations used in Table 27 include: PD, poorly differentiated; MD, moderately differentiated; NSCC, non-small cell carcinoma; SCC, squamous cell carcinoma; AC, adenocarcinoma; IDC, intvasive ductal carcinoma; ILC, invasive lobular carcinoma; Met, metastasis. Parenthesis indicates the tissue in which the tumor was found, if other than the tissue of origin.

TABLE 28


Spectrum	Tissue	Relative Expression

MPM51	Breast—IIC	1.29
MPM66	Breast—IIC	2.25
MPM67	Breast—IIC	1.80
MPM81	Breast—MetC	2.50
MPM50	Breast—MetC	1.24
MPM68	Breast—MetC	10.23
MPM70	Breast—MetC	10.22
MPM71	Breast—MetC	1.92

Expression array-based analysis of human 32144 mRNA expression in invasive indolent breast carcinomas (IIC) and metastatic breast carcinomas (MetC). 2/5 metastatic breast carcinomas displayed an elevated level of 32144 expression, while 0/3 invasive indolent breast carcinomas displayed an elevation in 32144 expression, suggesting a correlation between elevated 32144 expression and tumor metastasis.

Human 32235

The present invention is based, in part, on the discovery of a novel aminotransferase family member, referred to herein as “32235”.

The human 32235 sequence (SEQ ID NO:67), which is approximately 1816 nucleotides long including untranslated regions, contains a predicted methionine-initiated coding sequence of about 1350 nucleotides, not including the termination codon (nucleotides 84-1433 of SEQ ID NO:67; 1-1350 of SEQ ID NO:69). The coding sequence encodes a 450 amino acid protein (SEQ ID NO:68).

An alignment of the aminotransferase class III domain of human 32235 with a consensus amino acid sequence derived from a hidden Markov model (HMM) from PFAM shows the consensus amino acid sequence (SEQ ID NO:70) aligns with amino acids 23 to 437 of SEQ ID NO:68.

A BLAST alignment of a first region of the aminotransferase class III domain of human 32235 with a consensus amino acid sequence of a domain derived from the ProDomain database (“AMINOTRANSFERASE CG8745 CG11241 PHOSPHATE PYRIDOXAL AMINOTRANSFERASES PRECURSOR BETA-ALAAT BETA-ALANINE-PYRUVATE;” No. PD082189; ProDomain Release 2001.1) shows amino acid residues 1 to 159 of the amino acid PD082189 consensus sequence (SEQ ID NO:71) aligns the first region of the human 32235 sequence (amino acid residues 84 to 246 of SEQ ID NO:68).

A BLAST alignment of a second region of the aminotransferase class III domain of human 32235 with a consensus amino acid sequence of a domain derived from the ProDomain database (“AMINOTRANSFERASE PYRIDOXAL ADENOSYLMETHIONINE-8-AMINO-7-OXONONANOATE PHOSPHATE TRANSAMINASE BIOSYNTHESIS ACID DAPA 78-DIAMINO-PELARGONIC;” No. PD000519; ProDomain Release 2001. 1) shows amino acid residues 12 to 68 of the amino acid PD000519 consensus sequence (SEQ ID NO:72), while the upper amino acid sequence corresponds to the second region of the human 32235 sequence (amino acid residues 308 to 363 of SEQ ID NO:68).

A CLUSTAL W alignment of human 32235 with human and mouse beta-alanine pyruvate aminotransferase (Accession No. AR105920 in GenBank, and BAB28878 in GenPept, respectively) shows nucleotides 1 to 1844 of BAB28878 (SEQ ID NO:73) and nucleotides 1 to 1786 of AR105920 (SEQ ID NO:74) align with nucleotides 1 to 1816 of human 32235 (SEQ ID NO:69). CLUSTAL W (v 1.74; Thompson et al. (1994) Nuc. Acids Res. 22:4673-80) uses dynamically varied gap penalties for progressive sequence alignments.

A CLUSTAL W alignment of human 32235 with human ornithine aminotransferase (Accession No. P04181 in Swissprot) and human 4-aminobutyrate aminotransferase (Accession No. P80404 in Swissprot) shows amino acids 1 to 439 of P04181 (SEQ ID NO:75) aligns with amino acids 1 to 450 of human 32235 and amino acids 1 to 500 of P80404 (SEQ ID NO:76).

Human 32235 contains the following regions or other structural features (for general information regarding PFAM identifiers, PS prefix and PF prefix domain identification numbers, refer to Sonnhammer et al. (1997) Protein 28:405-420): an aminotransferase class III domain (PFAM Accession No. PF00202) located at about amino acid residues 23 to 437 of SEQ ID NO:68; one coiled coil structure (PSORT) located at about amino acids 416 to 446 of SEQ ID NO:68; one aminotransferase class III pyridoxal-phosphate attachment site (ProSite PS00600) located at about amino acids 203 to 206 of SEQ ID NO:68; three protein kinase C phosphorylation sites (ProSite PS00005) located at about amino acids 22 to 24, 173 to 175, and 445 to 447 of SEQ ID NO:68; six casein kinase II phosphorylation sites (ProSite PS00006) located at about amino acids 99 to 102, 112 to 115, 146 to 149, 199 to 202, 302 to 305, and 434 to 437 of SEQ ID NO:68; four N-myristoylation sites (ProSite PS00008) located at about amino acids 113 to 118, 241 to 246, 312 to 317, and 364 to 369 of SEQ ID NO:68; and one amidation site (ProSite PS00009) located at about amino acids 203 to 206 of SEQ ID NO:68.

A hydropathy plot of human 32235 was performed. Polypeptides of the invention include fragments which include: all or part of a hydrophobic sequence; all or part of a hydrophilic sequence; or a sequence which includes a cysteine residue.

The 32235 protein contains a significant number of structural characteristics in common with members of the aminotransferase family. As used herein, the term “aminotransferase” includes a protein or polypeptide which is capable of transferring an amino group from an amino acid to an oxo acid.

Members of the aminotransferase family of proteins are generally cytoplasmic or mitochondrial and play a pivotal role in the metabolism of amino acids. An alignment of the 32235 protein with human beta-alanine pyruvate aminotransferase (Accession No. in GenBank AR105920) demonstrates about 99% sequence identity between the two sequences (as calculated by CLUSTAL W). An alignment of the 32235 protein with a mouse ortholog of human beta-alanine pyruvate aminotransferase (Accession No. in GenPept BAB28878) demonstrates about 87% sequence identity between the two sequences (as calculated by CLUSTAL W).

A 32235 polypeptide can include an “aminotransferase class III domain” or regions homologous with an “aminotransferase class III domain”. A 32235 polypeptide can further include a “coiled coil structure” or regions homologous with a “coiled coil structure,” and at least one aminotransferase class III pyridoxal-phosphate attachment site.

As used herein, the term “aminotransferase class III domain” includes an amino acid sequence of about 400 to 500 amino acid residues in length and having a bit score for the alignment of the sequence to the aminotransferase class III domain (HMM) of at least 150. Preferably an aminotransferase class III domain mediates the transfer of an amino group from an amino acid to an oxo acid. Preferably, an aminotransferase class III domain includes at least about 400 to 500 amino acids, more preferably about 425 to 475 amino acid residues, or about 440 to 460 amino acids and has a bit score for the alignment of the sequence to the aminotransferase class III domain (HMM) of at least 150, more preferably at least 200, most preferably 250 or greater.

The aminotransferase class III domain can include a ProSite aminotransferase class III pyridoxal-phosphate attachment site (signature sequence ProSite PS00600), or sequences homologous thereto. The ProSite aminotransferase class III pyridoxal-phosphate attachment site has the following consensus sequence: [LIVMFYWC](2)-x-D-E-[IVA]-x(2)-G-[LIVMFAGC]-x(0,1)-[RSACLI]-x-[GSAD]-x(12,16)-D-[LIVMFC]-[LIVMFYSTA]-x(2)-[GSA]-K-x(3)-[GSTADNV]-[GSAC] (SEQ ID NO:77). In the above conserved signature sequence, and other motifs or signature sequences described herein, the standard IUPAC one-letter code for the amino acids is used. Each element in the pattern is separated by a dash (-); square brackets ([]) indicate the particular residues that are accepted at that position; x indicates that any residue is accepted at that position; and numbers in parentheses (( )) indicate the number of residues represented by the accompanying amino acid.

The aminotransferase class III domain preferably includes the following highly conserved residues and regions: a nucleotide binding region (amino acids 251 to 256 of SEQ ID NO:68); a glutamic acid residue that may interact with the 3′-OH of pyridoxal-5′-phosphate (E213 in SEQ ID NO:68); an aspartate residue that may interact with the N1 nitrogen of pyridoxal-5′-phosphate (D246 in SEQ ID NO:68); and a lysine residue that may form a Schiff base with pyridoxal-5′-phosphate (K278 in SEQ ID NO:68). In certain embodiments, the aminotransferase class III domain may also include the following conserved residues: G39, Y41, D44, G47, D52, S55, G61, V68, R83, G113, A120, P183, A208, G220, F243, E247, Q249, G251, G256, G283, T309, G312, P314, E330, L32, A336, G340, L343, L347, V360, R361, G362, G364, F411, and P413 in SEQ ID NO:2 that may play a catalytic and/or structural role.

The aminotransferase class III domain (HMM) has been assigned the PFAM Accession Number PF00202. The aminotransferase class III domain (amino acids 23 to 437 of SEQ ID NO:68) of human 32235 aligns with the PFAM aminotransferase class III domain consensus amino acid sequence (SEQ ID NO:70) derived from a hidden Markov model.

In a preferred embodiment, a 32235 polypeptide or protein has an “aminotransferase class III domain” or a region which includes at least about 400 to 500 amino acids, more preferably about 425 to 475 amino acid residues, or about 440 to 460 amino acid residues and has at least about 60%, 70%, 80%, 90%, 95%, 99%, or 100% homology with an “aminotransferase class III domain,” e.g., the aminotransferase class III domain of human 32235 (e.g., residues 23 to 437 of SEQ ID NO:68).

To identify the presence of an “aminotransferase class III domain” in a 32235 protein sequence, and make the determination that a polypeptide or protein of interest has a particular profile, the amino acid sequence of the protein can be searched against the Pfam database of HMMs (e.g., the Pfam database, release 2.1) using the default parameters. For example, the hmmsf program, which is available as part of the HMMER package of search programs, is a family specific default program for MILPAT0063 and a score of 15 is the default threshold score for determining a hit. Alternatively, the threshold score for determining a hit can be lowered (e.g., to 8 bits). A description of the Pfam database can be found in Sonhammer et al. (1997) Proteins 28:405-420 and a detailed description of HMMs can be found, for example, in Gribskov et al. (1990) Meth. Enzymol. 183:146-159; Gribskov et al. (1987) Proc. Natl. Acad. Sci. USA 84:4355-4358; Krogh et al. (1994) J. Mol. Biol. 235:1501-1-531; and Stultz et al. (1993) Protein Sci. 2:305-314, the contents of which are incorporated herein by reference. A search was performed against the HMM database resulting in the identification of an “aminotransferase class III domain” in the amino acid sequence of human 32235 at about residues 23 to 437 of SEQ ID NO:68.

For further identification of an “aminotransferase class III domain” in a 32235 protein sequence, and make the determination that a polypeptide or protein of interest has a particular profile, the amino acid sequence of the protein can be searched against a database of domains, e.g., the ProDom database (Corpet et al. (1999), Nucl. Acids Res. 27:263-267). The ProDom protein domain database consists of an automatic compilation of homologous domains. Current versions of ProDom are built using recursive PSI-BLAST searches (Altschul et al. (1997) Nucleic Acids Res. 25:3389-3402; Gouzy et al. (1999) Computers and Chemistry 23:333-340) of the SWISS-PROT 38 and TREMBL protein databases. The database automatically generates a consensus sequence for each domain. A BLAST search was performed against the HMM database resulting in the identification of a first and second region of an “aminotransferase class III domain” domain in the amino acid sequence of human 32235 at about residues 84 to 246 and 308 to 363 of SEQ ID NO:68.

A 32235 family member can include at least one amino transferase class III domain. A 32235 family member can further include a coiled coil structure and an aminotransferase class III pyridoxal-phosphate attachment site (ProSite PS00600). Furthermore, a 32235 family member can include at least one, two, preferably three protein kinase C phosphorylation sites (ProSite PS00005); at least one, two, three, four, five, preferably six casein kinase II phosphorylation sites (ProSite PS00006); at least one, two, three, and preferably four N-myristoylation sites (ProSite PS00008); and at least one amidation site (ProSite PS00009).

As the 32235 polypeptides of the invention can modulate 32235-mediated activities, they can be useful for developing novel diagnostic and therapeutic agents for aminotransferase-associated or other 32235-associated disorders, as described below.

As used herein, an “aminotransferase-associated activity” includes an activity which involves transfer of an amino group from an amino acid to an oxo acid. Members of the family can play a role in metabolic disorders, e.g., disorders of amino acid metabolism.

As used herein, a “32235 activity”, “biological activity of 32235” or “functional activity of 32235”, refers to an activity exerted by a 32235 protein, polypeptide or nucleic acid molecule on e.g., a 32235-responsive cell or on a 32235 substrate, e.g., a protein substrate, as determined in vivo or in vitro. In one embodiment, a 32235 activity is a direct activity, such as an association with a 32235 target molecule. A “target molecule” or “binding partner” is a molecule with which a 32235 protein binds or interacts in nature. In an exemplary embodiment, 32235 is an enzyme for a substrate, e.g., an amino acid substrate such as L-alanine or an oxo acid substrate such as pyruvate.

A 32235 activity can also be an indirect activity, e.g., a cellular signaling activity mediated by interaction of the 32235 protein with a 32235 receptor. Based on the above-described sequence structures and similarities to molecules of known function, the 32235 molecules of the present invention can have similar biological activities as aminotransferase family members. For example, the 32235 proteins of the present invention can have one or more of the following activities: (1) the ability to modulate metabolism, e.g., amino acid metabolism; (2) the ability to bind an amino acid, e.g., L-alanine; (3) the ability to bind an oxo acid, e.g., pyruvate; (4) the ability to bind a co-factor, e.g., pyridoxal-5′-phosphate; and (5) the ability to catalyze the transfer of an amino group from an amino acid to an oxo acid, e.g., from L-alanine to pyruvate.

The 32235 molecules of the invention can modulate the activities of cells in tissues where they are expressed. For example, 32235 mRNA is expressed in lung tumors, prostate tumors, ovarian tumors, colon tumors, breast tumors, normal artery, normal heart, heart under congestive heart failure, kidney, skeletal muscle, pancreas, normal brain hypothalamus, and nerve. Accordingly, the 32235 molecules of the invention can act as therapeutic or diagnostic agents for cellular proliferative, cardiovascular, renal, muscular, pancreatic, neurological disorders, and metabolic.

The 32235 molecules can be used to treat cellular proliferative and/or differentiative disorders in part because 32235 mRNA is expressed in tumor tissues, e.g., breast tumors, lung tumors, prostate tumors, ovarian tumors and colon tumors. Examples of cellular proliferative and/or differentiative disorders include cancer, e.g., carcinoma, sarcoma, metastatic disorders or hematopoietic neoplastic disorders, e.g., leukemias. A metastatic tumor can arise from a multitude of primary tumor types, including but not limited to those of prostate, colon, lung, breast and liver origin.

The 32235 molecules can be used to treat pancreatic disorders in part because 32235 mRNA is expressed in the pancreas.

The 32235 molecules can be used to treat endothelial cell disorders in part because 32235 mRNA is expressed in endothelial tissues, e.g., human umbilical vein endothelial cells (HUVEC) and human microvascular endothelial cells (HMVEC).

The 32235 molecules can be used to treat pain disorders because 32235 mRNA is expressed in neurological tissues, e.g., nerves and the hypothalamus.

Thus, the 32235 molecules can act as novel diagnostic targets and therapeutic agents for controlling one or more cellular proliferative, cardiovascular, renal, muscular, pancreatic, neurological or other aminotransferase disorder. As used herein, “aminotransferase disorders” are diseases or disorders whose pathogenesis is caused by, is related to, or is associated with aberrant or deficient aminotransferase protein function or expression. Examples of such disorders, e.g., aminotransferase-associated or other 32235-associated disorders, include but are not limited to metabolic disorders.

The 32235 molecules can be used to treat metabolic disorders in part because aberrant or deficient function or expression of aminotransferase family members results in the inability to fully degrade essential amino acids. Diseases of metabolic imbalance include, but are not limited to, obesity, anorexia nervosa, cachexia, lipid disorders, and diabetes.

Gene Expression Analysis of 32235

Human 32235 expression was measured by TaqMan® quantitative PCR (Perkin Elmer Applied Biosystems) in cDNA prepared from a variety of normal and diseased (e.g., cancerous) human tissues or cell lines.

The results indicate significant 32235 expression in tumors, cardiovascular, renal, muscular, pancreatic, and neurological tissues.

Tables

32235 was first identified from a T×P experiment which profiled three distinct ovarian carcinoma cell lines that were grown on plastic, soft agar, and as subcutaneous xenograft tumors (see Table 29). 32235 was found to be upregulated when the cells were grown either on soft agar or as xenograft tumors compared to growth on plastic.

TABLE 29


TxP analysis

	Cell line	32235 expression

	HEY (plastic)	1.4841
	HEY (soft agar)	1.6636
	SKOV-3 #1 (plastic)	1.0752
	SKOV-3 #1 (soft agar)	1.4575
	SKOV-3 #1 (tumor)	1.7173
	SKOV-3 #2 (plastic)	1.4160
	SKOV-3 #2 (soft agar)	1.9042
	SKOV-3 #2 (tumor)	2.1054
	SKOV-3 variant #1 (plastic)	1.2800
	SKOV-3 variant #1 (soft agar)	1.4000
	SKOV-3 variant #1 (tumor)	1.9748
	SKOV-3 variant #2 (plastic)	1.4500
	SKOV-3 variant #2 (soft agar)	1.3626
	SKOV-3 variant #2 (tumor)	1.5179

The expression of 32235 was also increased with addition of the growth factor EGF to serum free culture media of the SKOV-3 cell line for 15, 30, or 60 minutes (see Table 30). Clinical data comparing expression of 32235 in isolated ovarian epithelial cells vs. ascites (see Table 31), across a range of tissues (see Table 32), and expression in normal and diseased tissues (see Table 33), all indicate that 32235 is upregulated in tumor tissues compared to normal tissues. 32235 is also expressed in several xenograft friendly cell lines (see Table 34).

TABLE 30


TaqMan ® analysis of the
ovarian carcinoma cell line SKOV-3 ± EGF

	Cell line	Relative 32235 expression

	SKOV-3 (without EGF)	4.2
	SKOV-3 (with EGF 15′)	5.8
	SKOV-3 (with EGF 30′)	5.0
	SKOV-3 (with EGF 60′)	5.8

TABLE 31


TaqMan ® analysis of clinical human isolated
ovarian epithelial cells compared to clinical ovarian ascites

		Relative 32235
	Tissue	expression

	Ovary (normal)	0.9
	Ovary (normal)	0.5
	Ovary (ascites)	1.1
	Ovary (ascites)	1.6

TABLE 32


TaqMan ® organ recital

		Relative 32235
	Tissue	expression

	Artery (normal)	22.9
	Aorta (diseased)	10.7
	Vein (normal)	4.0
	Coronary smooth muscle	11.6
	HUVEC¹	28.6
	Hemangioma	11.4
	Heart (normal)	17.1
	Heart (CHF²)	18.3
	Kidney	17.9
	Skeletal muscle	24.0
	Adipose (normal)	4.8
	Pancreas	15.8
	Primary osteoblasts	5.5
	Osteoclasts (differentiated)	0.9
	Skin (normal)	7.0
	Spinal cord (normal)	8.8
	Brain hypothalamus (normal)	19.2
	Nerve	27.6
	Dorsal root ganglion	11.1
	Breast (normal)	10.9
	Breast (tumor)	6.6
	Ovary (normal)	13.5
	Ovary (tumor)	2.8
	Prostate (normal)	11.5
	Prostate (tumor)	15.6
	Salivary glands	2.7
	Colon (normal)	4.7
	Colon (tumor)	10.3
	Lung (normal)	3.0
	Lung (tumor)	10.4
	Lung (COPD³)	11.6
	Colon (IBD⁴)	2.8
	Liver (normal)	8.5
	Liver (fibrosis)	11.2
	Spleen (normal)	5.4
	Tonsil (normal)	8.1
	Lymph node (normal)	6.3
	Small intestine (normal)	3.5
	Macrophages	0.5
	Synovium	1.8
	Bone marrow MNC⁵	4.4
	Activated peripheral blood MNC⁵	1.9
	Neutrophils	5.3
	Megakaryocytes	8.9
	Erythroid	11.7

TABLE 33


TaqMan ® analysis comparing clinical tumors
with their normal tissue counterparts

Source	Tissue	Relative 32235 expression

PIT 400	Breast (normal)	32.7
PIT 372	Breast (normal)	18.8
CHT 559	Breast (normal)	0.7
CLN 168	Breast (tumor, IDC¹)	5.5
MDA 304	Breast (tumor, MD-IDC²)	3.1
CHT 2002	Breast (tumor, IDC¹)	8.3
CHT 562	Breast (tumor, IDC¹)	4.0
NDR 138	Breast (tumor, ILC³)	10.2
CHT 1841	Lymph node (breast met.)	18.9
PIT 58	Lung (breast met.)	5.0
CHT 620	Ovary (normal)	6.7
PIT 208	Ovary (normal)	11.2
CLN 012	Ovary (tumor)	10.8
CLN 07	Ovary (tumor)	2.9
CLN 17	Ovary (tumor)	12.7
MDA 25	Ovary (tumor)	24.3
MDA 216	Ovary (tumor)	2.9
PIT 298	Lung (normal)	4.4
MDA 185	Lung (normal)	6.3
CLN 930	Lung (normal)	6.6
MPI 215	Lung (tumor, SmC⁴)	7.3
MDA 259	Lung (tumor, PD-NSCC⁵)	23.9
CHT 832	Lung (tumor, PD-NSCC⁵)	6.4
MDA 262	Lung (tumor, SCC⁶)	9.6
CHT 793	Lung (tumor, ACA⁷)	4.6
CHT 331	Lung (tumor, ACA⁷)	19.0
CHT 405	Colon (normal)	9.4
CHT 523	Colon (normal)	10.7
CHT 371	Colon (normal)	6.5
CHT 382	Colon (tumor, MD⁸)	7.0
CHT 528	Colon (tumor, MD⁸)	11.8
CLN 609	Colon (tumor)	11.2
NDR 210	Colon (tumor, PD⁹)	36.4
CHT 340	Colon (liver met.)	26.8
NDR 100	Colon (liver met.)	15.8
PIT 260	Liver (normal, female)	6.3
CHT 1653	Cervix (SCC⁶)	8.5
CHT 569	Cervix (SCC⁶)	1.7
A24	HMVEC¹⁰(arrested)	10.0
C48	HMVEC¹⁰(proliferating)	14.7
Pooled	Hemangiomas	3.6
HCT 116 N22	Normoxic	30.0
HCT 116 H22	Hypoxic	14.4

TABLE 34


TaqMan ® analysis of xenograft friendly cell lines

Cell line	Tissue	Relative 32235 expression

MCF-7	Breast (tumor)	115.8
ZR75	Breast (tumor)	62.9
T47D	Breast (tumor)	40.4
MDA231	Breast (tumor)	16.9
MDA435	Breast (tumor)	20.5
SKBr3	Breast (tumor)	31.8
DLD1	Colon (tumor, stage C)	397.8
SW480	Colon (tumor, stage B)	42.2
HCT116	Colon (tumor)	45.6
HT29	Colon (tumor)	27.5
Colo 205	Colon (tumor)	65.4
NCIH125		69.8
NCIH67		44.0
NCIH322		44.5
NCIH460		56.1
A549	Lung (tumor)	122.9
NHBE¹	Lung	60.2
SKOV-3	Ovary (tumor)	11.8
OVCAR-3	Ovary (tumor)	22.7
293	Baby kidney	116.2
293T	Baby kidney	289.2

In situ hybridization (ISH) localized 32235 to the epithelial tumor component of 7/8 ovarian tumors and 3/3 lung tumors. No expression was found in normal ovarian surface epithelium (see Table 35).

TABLE 35


In situ hybridization

	Tissue	Diagnosis	Results

Ovary: 7/8 Tumors; 1/1 Met; 0/2 Normals

Ovary T	Endometrial adenocarcinoma	(−/−)
Ovary T	Endometrial adenocarcinoma	(+++/+)
Ovary T	Endometrial adenocarcinoma	(+/+)
Ovary T	PD-Serous	(+++/+)
Ovary T	MD-Adenocarcinoma	(+/+)
Ovary T	Serous carcinoma	(++/+)
Ovary T	PD-Serous carcinoma	(++/+)
Ovary T	PD-Clear cell	(++/+)
Ovary M	Ovarian met	(+++/+)
Ovary N	Normal	(−/−)
Ovary N	Normal ovarian stroma	(−/−)

Lung: 3/3 Tumors

Lung T	Non-small	(++/+)
Lung T	Small cell	(++/+)
Lung T	Small cell	(+/+)

Human 23565

The present invention is based, in part, on the discovery of a novel zinc carboxypeptidase family member, referred to herein as “23565”.

The human 23565 sequence (SEQ ID NO:78), which is approximately 1687 nucleotides long including untranslated regions, contains a predicted methionine-initiated coding sequence of about 1308 nucleotides, (nucleotides 160-1467 of SEQ ID NO:78, 1-1308 of SEQ ID NO:80) not including the termination codon. The coding sequence encodes a 436 amino acid protein (SEQ ID NO:79).

The human 23565 protein of SEQ ID NO:79 includes an amino-terminal hydrophobic amino acid sequence, consistent with a signal sequence, of about 34 amino acids (from amino acid 1 to about amino acid 34 of SEQ ID NO:79), which upon cleavage results in the production of a mature protein form. The mature protein form is approximately 402 amino acid residues in length (from about amino acid 35 to amino acid 436 of SEQ ID NO:79).

An alignment of the zinc carboxypeptidase domain of human 23565 with a consensus amino acid sequence derived from a hidden Markov model (HMM) from PFAM shows the consensus amino acid sequence (SEQ ID NO:81) aligns with amino acids 139 to 419 of SEQ ID NO:79.

An alignment of the zinc carboxypeptidase domain of human 23565 with a consensus amino acid sequence derived from a hidden Markov model (HMM) from SMART shows the consensus amino acid sequence (SEQ ID NO:82) aligns with,amino acids 139 to 419 of SEQ ID NO:79.

An alignment of the carboxypeptidase activation peptide of human 23565 with a consensus amino acid sequence derived from a hidden Markov model (HMM) from PFAM shows the consensus amino acid sequence (SEQ ID NO:83), while the lower amino acid sequence corresponds to amino acids 41 to 118 of SEQ ID NO:79.

Human 23565 contains the following regions or other structural features: one zinc carboxypeptidase domain (PFAM Accession Number PF00246) located at about amino acid residues 139 to 419 of SEQ ID NO:79, which includes one predicted zinc carboxypeptidase zinc-binding region 1 signature from about amino acid residues 187 to 209 of SEQ ID NO:79; and one predicted zinc carboxypeptidase zinc-binding region 2 signature from about amino acid residues 323 to 333 of SEQ ID NO:79; one carboxypeptidase activation peptide (PFAM Accession Number PF02244) located at about amino acid residues 41 to 118 of SEQ ID NO:79; one signal peptide located at about amino acids 1 to 34 of SEQ ID NO:79; four N-Glycosylation sites (PS00001) at about amino acids 36 to 39, 171 to 174, 256 to 259, and 281 to 284 of SEQ ID NO:79; one Glycosaminoglycan attachment site (PS00002) at about amino acid 276 to 279 of SEQ ID NO:79; two Protein Kinase C phosphorylation sites (PS00005) at about amino acids 124 to 126, and 258 to 260 of SEQ ID NO:79; seven Casein Kinase II phosphorylation sites (PS00006) at about amino 14 to 17, 141 to 144, 147 to 150, 238 to 241, 299 to 302, 416 to 419, and 426 to 429 of SEQ ID NO:79; one Tyrosine kinase phosphorylation site (PS00007) at about amino acid 351 to 358 of SEQ ID NO:79; and eight N-myristylation sites (PS00008) at about amino acid 3 to 8, 100 to 105, 206 to 211, 267 to 272, 279 to 284, 314 to 319, 368 to 373, and 389 to 394 of SEQ ID NO:79.

For general information regarding PFAM identifiers, PS prefix and PF prefix domain identification numbers, refer to Sonnhammer et al. (1997) Protein 28: 405-420.

A hydropathy plot of human 23565 was performed. Polypeptides of the invention include fragments which include: all or part of a hydrophobic sequence, e.g., the sequence from about amino acid 105 to 119, from about 222 to 235, from about 260 to 267 and from about 200 to 310 of SEQ ID NO:79; all or part of a hydrophilic sequence, e.g., the sequence of from about amino acid 115 to 142, from about 245 to 258, and from about 280 to 301 of SEQ ID NO:79; a sequence which includes a Cys, or a glycosylation site.

The 23565 protein contains a significant number of structural characteristics in common with members of the zinc carboxypeptidase family. The zinc carboxypeptidase family of proteins are structurally and functionally related, and are characterized by the following signature patterns: a zinc carboxypeptidase zinc-binding region 1 signature [PK]-x-[LIVMFY]-x-[LIVMFY]-x(4)-H-[STAG]-x-E-x-[LIVM]-[STAG]-x(6)-[LIVMFYTA] (SEQ ID NO:84), wherein H and E are zinc ligands, and a zinc carboxypeptidase zinc-binding region 2 signature H-[STAG]-x(3)-[LIVME]-x(2)-[LIVMFYW]-P-[FYW] (SEQ ID NO:85), wherein H is a zinc ligand. 23565, a member of the carboxypeptidase family shows good homology with critical residues of known family members. A 23565 protein typically contains one or more sequences that conform to each of the signature patterns. For example, a 23565 protein contains the sequence PAIWIDTGHSREWITHATGIWT (SEQ ID NO:86) located at amino acids 187 to 209 of SEQ ID NO:79, which corresponds to the zinc carboxypeptidase zinc-binding region 1 signature. A 23565 protein can also include the sequence HSYSQMLMYPY (SEQ ID NO:87) located at amino acids 323 to 333 of SEQ ID NO:79, which corresponds to the zinc carboxypeptidase zinc-binding region 2 signature. Carboxypeptidases are known to degrade peptide hormone and growth factors.

A 23565 polypeptide can include a “zinc carboxypeptidase domain” or regions homologous with a “zinc carboxypeptidase domain”. As used herein, the term “zinc carboxypeptidase domain” includes an amino acid sequence of about 100 to 400 amino acid residues in length and having a bit score for the alignment of the sequence to the zinc carboxypeptidase domain (HMM) of at least 200. Preferably, a zinc carboxypeptidase domain includes at least about 200 to 350 amino acids, more preferably about 250 to 300 amino acid residues, or about 275 to 285 amino acids and has a bit score for the alignment of the sequence to the zinc carboxypeptidase domain (HMM) of at least 250, 300, 350, 400 or greater. In one embodiment, a zinc carboxypeptidase domain includes one zinc carboxypeptidase zinc-binding region 1 signature: PAIWIDTGHSREWITHATGIWT (SEQ ID NO:86) located at amino acids 187 to 209 of SEQ ID NO:79, wherein the H and E residues are zinc ligands; and one zinc carboxypeptidase zinc-binding region 2 signature: HSYSQMLMYPY (SEQ ID NO:87) located at amino acids 323 to 333 of SEQ ID NO:79, wherein the H is a zinc ligand. The zinc carboxypeptidase domain (HMM) has been assigned the PFAM Accession Number PF00246. The zinc carboxypeptidase domain (HMM) has also been assigned the SMART identifier zn_carb. The zinc carboxypeptidase domain (amino acids 139 to 419 of SEQ ID NO:79) of human 23565 aligns with a consensus amino acid sequence (SEQ ID NOs: 81 and 82) derived from a hidden Markov model.

In a preferred embodiment 23565 polypeptide or protein has a “zinc carboxypeptidase domain” or a region which includes at least about 200 to 350 more preferably about 250 to 300, or 275 to 285 amino acid residues and has at least about 60%, 70% 80% 90% 95%, 99%, or 100% homology with a “zinc carboxypeptidase domain,” e.g., the zinc carboxypeptidase domain of human 23565 (e.g., residues 139 to 419 of SEQ ID NO:79).

The zinc carboxypeptidase family member may also include a carboxypeptidase activation peptide, which is a pro-segment motif accounting for up to about a quarter of the total length of the peptidase and responsible for modulation of folding and activity of the enzyme. Preferably, the carboxypeptidase activation peptide includes at least about 20 to 200 amino acids, more preferably about 50 to 100 amino acid residues, or about 70 to 80 amino acids and has a bit score for the alignment of the sequence to the carboxypeptidase activation peptide (HMM) of at least 50, 70, 90, 100, or greater. The. carboxypeptidase activation peptide motif has been assigned the PFAM Accession Number PF02244. The zinc carboxypeptidase domain (amino acids 41 to 118 of SEQ ID NO:79) of human 23565 aligns with a consensus amino acid sequence derived from a hidden Markov model.

In a preferred embodiment 23565 polypeptide or protein has a “carboxypeptidase activation peptide” or a region which includes at least about 20 to 200 more preferably about 50 to 100 or 70 to 80 amino acid residues and has at least about 50%, 60%, 70% 80% 90% 95%, 99%, or 100% homology with a “carboxypeptidase activation peptide,” e.g., the carboxypeptidase activation peptide of human 23565 (e.g., residues 41 to 118 of SEQ ID NO:79).

To identify the presence of a “zinc carboxypeptidase” domain or a “carboxypeptidase activation peptide” in a 23565 protein sequence, and make the determination that a polypeptide or protein of interest has a particular profile, the amino acid sequence of the protein can be searched against the Pfam database of HMMs (e.g., the Pfam database, release 2.1) using the default parameters. For example, the hmmsf program, which is available as part of the HMMER package of search programs, is a family specific default program for MILPAT0063 and a score of 15 is the default threshold score for determining a hit. Alternatively, the threshold score for determining a hit can be lowered (e.g., to 8 bits). A description of the Pfam database can be found in Sonhammer et al. (1997) Proteins 28(3): 405-420 and a detailed description of HMMs can be found, for example, in Gribskov et al.(1990) Meth. Enzymol. 183:146-159; Gribskov et al. (1987) Proc. Natl. Acad. Sci. USA 84:4355-4358; Krogh et al.(1994) J. Mol. Biol. 235:1501-1531; and Stultz et al.(1993) Protein Sci. 2:305-314, the contents of which are incorporated herein by reference. A search was performed against the HMM database resulting in the identification of a “zinc carboxypeptidase” domain in the amino acid sequence of human 23565 at about residues 139 to 419 of SEQ ID NO:79, and the identification of a “carboxypeptidase activation peptide” in the amino acid sequence of human 23565 at about residues 41 to 118 of SEQ ID NO:79.

To identify the presence of a “zinc carboxypeptidase” domain in a 23565 protein sequence, and make the determination that a polypeptide or protein of interest has a particular profile, the amino acid sequence of the protein can be searched against a SMART database (Simple Modular Architecture Research Tool) of HMMs as described in Schultz et al. (1998), Proc. Natl. Acad. Sci. USA 95: 5857 and Schultz et al. (200) Nucl. Acids Res 28:231. The database contains domains identified by profiling with the hidden Markov models of the HMMer2 search program (R. Durbin et al. (1998) Biological sequence analysis: probabilistic models of proteins and nucleic acids. Cambridge University Press). The database also is extensively annotated and monitored by experts to enhance accuracy. A search was performed against the HMM database resulting in the identification of a “zinc carboxypeptidase” domain in the amino acid sequence of human 23565 at about residues 139 to 419 of SEQ ID NO:79.

In yet another embodiment, the 23565 molecule can further include a signal sequence. As used herein, a “signal sequence” refers to a peptide of about 20 to 50 amino acid residues in length which occurs at the N-terminus of secretory and integral membrane proteins and which contains a majority of hydrophobic amino acid residues. For example, a signal sequence contains at least about 30 to 40 amino acid residues, preferably about 34 amino acid residues, and has at least about 40-70%, preferably about 50-65%, and more preferably about 55-60% hydrophobic amino acid residues (e.g., alanine, valine, leucine, isoleucine, phenylalanine, tyrosine, tryptophan, or proline). Such a “signal sequence”, also referred to in the art as a “signal peptide”, serves to direct a protein containing such a sequence to a lipid bilayer. For example, in one embodiment, a 23565 protein contains a signal sequence of about amino acids 1 to 34 of SEQ ID NO:79. The “signal sequence” is cleaved during processing of the mature protein. The mature 23565 protein corresponds to amino acids 35 to 436 of SEQ ID NO:79.

A 23565 polypeptide can optionally include at least one, two, preferably three N-glycosylation sites; at least one glycosaminoglycan attachment site; at least one, preferably two protein kinase C phosphorylation sites; at least one, two, three, four, five, six, preferably seven casein kinase II phosphorylation sites; at least one tyrosine kinase phosphorylation sites; and at least one, two, three, four, five, six, seven, preferably eight N-myristylation sites.

As the 23565 polypeptides of the invention may modulate 23565-mediated activities, they may be useful as of for developing novel diagnostic and therapeutic agents for 23565-mediated or related disorders, as described below.

As used herein, a “23565 activity,” “biological activity of 23565” or “functional activity of 23565,” refers to an activity exerted by a 23565 protein, polypeptide or nucleic acid molecule. For example, a 23565 activity can be an activity exerted by 23565 in a physiological milieu on, e.g., a 23565-responsive cell or on a 23565 substrate, e.g., a protein substrate. A 23565 activity can be determined in vivo or in vitro. In one embodiment, a 23565 activity is a direct activity, such as an association with a 23565 target molecule. A “target molecule” or “binding partner” is a molecule with which a 23565 protein binds or interacts in nature.

In an exemplary embodiment, 23565 is an enzyme for a polypeptide substrate.

A 23565 activity can also be an indirect activity, e.g., a cellular signaling activity mediated by interaction of the 23565 protein with a 23565 receptor. The features of the 23565 molecules of the present invention can provide similar biological activities as zinc carboxypeptidase family members. For example, the 23565 proteins of the present invention can have one or more of the following activities: (1) formation of a zinc ion complex with a carbonyl group of a substrate polypeptide and polarization of the carbon-oxygen bond; (2) formation of a tetrahedral intermediate due to attack of the carbonyl carbon by water in a reaction assisted by a carboxylate side chain of glutamate; (3) production of a dianion intermediate by rapid ionization of the tetrahedral intermediate produced; (4) cleavage of the C—N bond of the substrate to collapse the tetrahedral intermediate; (5) binding the carboxy-terminus of polypeptides; (6) hydrolyzing polypeptides to remove/release a carboxy-terminal residue; (7) participating in digestion of polypeptides/proteins; (8) processing prohormones; (9) regulating growth hormones; (10) modulating (e.g., stimulate) cell differentiation or proliferation, e.g., differentiation or proliferation of hematopoietic cells; (11) modulating hematopoiesis, e.g., erythropoiesis; (12) modulating apoptosis, of a cell, e.g., increase apoptosis of a cancer cell, e.g., a leukemic cell, (e.g., an erythroleukemia cell); or suppress apoptosis of a blood or erythroid cell; or (13) modulating transcriptional activity, e.g., cytokine transcriptional activity.

Taqman analysis revealed high levels of expression of 23565 mRNA in erythroid (GPA+) and megakaryocyte (CD61+) lineages in vivo, and in vitro, high levels of expression only during late megakaryocyte differentiation, low levels of expression in most tissues, and moderate expression in skeletal muscle and pituitary (Tables 36-40). Table 36 shows 23565 mRNA expression as determined by TaqMan assays in a panel of human tissues, including artery normal, aorta diseased, vein normal, coronary SMC, Human Umbilical Vein Endothelial Cells (HUVEC), heart, pancreas, skin, spinal cord, brain, adrenal glands, dorsal root gland (DRG), nerve, breast, ovary, colon, lung, liver, megakaryocytes, and erythroid. The highest 23565 mRNA expression was observed in megakaryocytes, followed by skeletal muscle, lymphnode, tonsil, and pituitary gland. Its expression is further enhanced in the erythroid lineage and increases as blood cell differentiation proceeds. Tables 37-40 show relative 23565 mRNA expression as determined by TaqMan assays on mRNA most derived from human hematological samples, e.g., bone marrow (BM), erythroid cells (Eryth), megakaryocytes (Meg), neutrophils (Neut), or a negative reference sample (NTC). 23565 mRNA was highly expressed in pooled megakaryocytes, glycophorin A (GPA) expressing cells, and BM CD 61+ cell. In Table 39, mRNA expression was detected at the indicated times in culture (e.g., 24 hrs., 48 hrs., days in culture). High levels of 23565 mRNA expression were observed in one sample of erythroid cells, especially day 7 (erythroid burst forming units (BFU) Eryth D7). In Table 40, high levels of 23565 mRNA expressions were observed in two samples of megakaryocyte cells, especially day 6 and day 10. This pattern of expression suggests a role for 23565 in the regulation of cytokine signaling during the development of cells of the erythroid lineage. Thus, inhibition of 23565 expression is expected to accelerate megakaryopoiesis by inhibiting degradation of growth factors critical for megakaryocyte growth. Accordingly, the 23565 molecules can act as novel diagnostic targets and therapeutic agents for controlling hematopoietic disorders.

As used herein, a “CD61-positive cell” or a “CD61-expressing cell” refers to a cell that expresses detectable levels of the CD61 antigen, preferably human CD61 antigen. CD61 recognizes a Mr 110-kilodalton (kDa) protein, also known as gpIIIa, the common β-subunit (integrin β3-chain) of the gpIIb/IIIa complex and the vitronectin receptor. The CD61 antigen is typically present on hematopoietic cells and hematopoietic colony-forming cells in the bone marrow.

As the 23565 polypeptides of the invention may modulate 23565-mediated activities, they may be useful as of for developing novel diagnostic and therapeutic agents for 23565-mediated or related disorders, e.g., blood cell- (e.g., erythroid-) associated disorders and other hematopoietic disorders.

Agents that modulate 23565 polypeptide or nucleic acid activity or expression can be used to treat anemias, in particular, drug-induced anemias or anemias associated with cancer chemotherapy, chronic renal failure, malignancies, adult and juvenile rheumatoid arthritis, disorders of hemoglobin synthesis, prematurity, and zidovudine treatment of HIV infection. A subject receiving the treatment can be additionally treated with a second agent, e.g., erythropoietin, to further ameliorate the condition.

As used herein, the term “erythropoietin” or “EPO” refers to a glycoprotein produced in the kidney, which is the principal hormone responsible for stimulating red blood cell production (erythrogenesis). EPO stimulates the division and differentiation of committed erythroid progenitors in the bone marrow. Normal plasma erythropoietin levels range from 0.01 to 0.03 Units/mL, and can increase up to 100 to 1,000-fold during hypoxia or anemia. Graber and Krantz, Ann. Rev. Med. 29: 51 (1978); Eschbach and Adamson, Kidney Intl. 28:1 (1985). Recombinant human erythropoietin (rHuEpo or epoietin alpha) is commercially available as EPOGEN.RTM. (epoietin alpha, recombinant human erythropoietin) (Amgen Inc., Thousand Oaks, Calif.) and as PROCRIT.RTM. (epoietin alpha, recombinant human erythropoietin) (Ortho Biotech Inc., Raritan, N.J.).

Aberrant expression or activity of the 23565 molecules may be involved in neoplastic disorders. Accordingly, treatment, prevention and diagnosis of cancer or neoplastic disorders related to hematopoietic cells and, in particular, cells of the erythroid lineage are also included in the present invention.

The 23565 nucleic acid and protein of the invention can also be used to treat and/or diagnose a variety of immune disorders.

The molecules of the invention may also modulate the activity of tissues in which they are expressed, e.g., skeletal muscle or pituitary, as well as other neoplastic tissues. For example, increase expression of 23565 molecules is detected on lung tumors compared to the normal lung. Accordingly, the 23565 molecules can act as novel diagnostic targets and therapeutic agents for controlling one or more of cellular proliferative and/or differentiative disorders.

Tissue Distribution of 23565 mRNA by TagMan Analysis

Endogenous human 23565 gene expression was determined using the Perkin-Elmer/ABI 7700 Sequence Detection System which employs TaqMan technology. Tissues tested include the human tissues and several cell lines shown in Tables 36-40.

TABLE 36


23565 mRNA Expression

Tissue Type	Mean	β 2 Mean	∂∂ Ct	Expression

Artery normal	40.0	20.1	19.9	0.0
Aorta diseased	39.9	22.9	17.0	0.0
Vein normal	40.0	20.2	19.8	0.0
Coronary SMC	38.7	19.7	18.9	0.0
HUVEC	40.0	21.2	18.9	0.0
Hemangioma	40.0	20.1	19.9	0.0
Heart normal	38.6	19.5	19.1	0.0
Heart CHF	40.0	21.3	18.7	0.0
Kidney	37.0	20.7	16.3	0.0
Skeletal Muscle	33.8	22.2	11.6	0.3
Liver normal	40.0	19.7	20.3	0.0
Small intestine normal	40.0	20.5	19.5	0.0
Adipose normal	36.7	19.2	17.5	0.0
Pancreas	35.8	22.2	13.6	0.0
primary osteoblast	38.5	19.8	18.7	0.0
Bladder	40.0	19.4	20.6	0.0
Adrenal Gland normal	36.0	19.4	16.6	0.0
Pituitary Gland normal	32.7	20.2	12.6	0.2
Spinal cord normal	38.7	22.0	16.6	0.0
Brain Cortex normal	36.2	22.2	14.0	0.0
Brain Hypothalamus normal	38.1	21.2	16.8	0.0
Nerve	40.0	20.9	19.1	0.0
DRG (Dorsal Root Ganglion)	39.4	21.6	17.9	0.0
Breast normal	34.8	20.3	14.5	0.0
Breast Tumor	33.9	19.6	14.3	0.0
Ovary normal	39.3	20.2	19.1	0.0
Ovary Tumor	37.1	19.9	17.2	0.0
Prostate BPH	37.5	20.1	17.4	0.0
Prostate Tumor	39.2	20.6	18.6	0.0
Colon normal	39.2	19.6	19.6	0.0
Colon Tumor	37.5	20.2	17.4	0.0
Lung normal	38.9	18.1	20.9	0.0
Lung tumor	38.6	20.2	18.4	0.0
Lung COPD	37.6	19.4	18.2	0.0
Colon IBD	40.0	20.5	19.5	0.0
Synovium	38.1	19.6	18.6	0.0
Tonsil normal	30.5	18.6	11.9	0.3
Lymph node normal	32.3	20.4	11.8	0.3
Liver fibrosis	38.4	21.2	17.3	0.0
Spleen normal	34.8	18.2	16.6	0.0
Macrophages	40.0	17.1	22.9	0.0
Progenitors	35.7	19.5	16.2	0.0
(erythroid, megakaryocyte,
neutrophil)
Megakaryocytes	28.3	19.3	9.1	1.8
Activated PBMC	37.0	18.1	18.9	0.0
Neutrophils	40.0	18.6	21.4	0.0
Erythroid	40.0	21.0	19.1	0.0
positive control	27.5	21.5	6.0	15.2

TABLE 37


23565 mRNA expression

		Avg	Avg			Rel exp
23565	Beta	23565	Beta	ΔCT	ΔΔCT	Rel exp

Heart PT 262	40	20.22	40.0	20.2	19.8	19.8	0.0
Brain MCL 400	33.83	20.49	33.8	20.5	13.3	13.3	0.1
Lung CHT 330	32.69	17.7	32.7	17.7	15.0	15.0	0.0
Liver NDR 379	35.89	21.6	35.9	21.6	14.3	14.3	0.0
Spleen 38O	31.04	19.22	31.0	19.2	11.8	11.8	0.3
Kidney 27	31.13	19.96	31.1	20.0	11.2	11.2	0.4
CD3 4 hr Rest LF164	30.31	18.56	30.3	18.6	11.8	11.8	0.3
CD3 4 hr Act LF164	31.72	19.49	31.7	19.5	12.2	12.2	0.2
CD3 24 hr Rest LF164	29.13	18.43	29.1	18.4	10.7	10.7	0.6
CD3 24 hr Act LF164	29.12	17.59	29.1	17.6	11.5	11.5	0.3
CD4 4 hr Rest LF164	33.04	20.24	33.0	20.2	12.8	12.8	0.1
CD4 4 hr Act LF164	31.06	18.77	31.1	18.8	12.3	12.3	0.2
CD4 24 hr Rest LF164	32.13	20.07	32.1	20.1	12.1	12.1	0.2
CD4 24 hr Act LF164	30.61	18.01	30.6	18.0	12.6	12.6	0.2
CD8 4 hr Rest LF164	28.92	18.7	28.9	18.7	10.2	10.2	0.8
CD8 4 hr Act LF164	32.51	20.5	32.5	20.5	12.0	12.0	0.2
CD8 24 hr Rest LF164	28.93	18.59	28.9	18.6	10.3	10.3	0.8
CD8 24 hr Act LF164	36.01	23.11	36.0	23.1	12.9	12.9	0.1
CD14-/19+ LF136	40	20.08	40.0	20.1	19.9	19.9	0.0
CD14 LF129	37.82	18.91	37.8	18.9	18.9	18.9	0.0
mBM CD14-/11b-/15+ LF120	34.97	19.25	35.0	19.3	15.7	15.7	0.0
mBM MNC LP7	37.55	18.97	37.6	19.0	18.6	18.6	0.0
mBM CD34+ LP92	35.16	20.25	35.2	20.3	14.9	14.9	0.0
BM CD34+ LF75	32.97	19.31	33.0	19.3	13.7	13.7	0.1
Cord Blood CD34+ LF101	34.37	19.86	34.4	19.9	14.5	14.5	0.0
GPA Hi LF156	29.57	20.25	29.6	20.3	9.3	9.3	1.6
Pooled Neut D6	32.24	19.17	32.2	19.2	13.1	13.1	0.1
Pooled Neut D10/12	36.43	18.97	36.4	19.0	17.5	17.5	0.0
Pooled Eryth D10/12	37.32	21.54	37.3	21.5	15.8	15.8	0.0
Pooled Meg D10/12	27.46	19.58	27.5	19.6	7.9	7.9	4.2
BM CD14-/15+ LF32	34.4	18.3	34.4	18.3	16.1	16.1	0.0
Grans LF157	37.43	17.11	37.4	17.1	20.3	20.3	0.0
K562	33.12	22.29	33.1	22.3	10.8	10.8	0.5
HL60	32.7	20.12	32.7	20.1	12.6	12.6	0.2
MF11 Stromal D32 post irrad	35.7	18.12	35.7	18.1	17.6	17.6	0.0
MF12 Stromal cntl	31.17	16.94	31.2	16.9	14.2	14.2	0.1
MF13 Stromal D2 post irrad	32.31	17.36	32.3	17.4	15.0	15.0	0.0
NTC	40	40	40.0	40.0	0.0	0.0

TABLE 38


23565 mRNA expression

		AVG	AVG			Rel exp
23565	Beta	23565	Beta	ΔCT	ΔΔCT	Rel exp

Lung CHT 330	33.82	18.22	33.8	18.2	15.6	15.6	0.0
Heart PT 262	37.86	20.47	37.9	20.5	17.4	17.4	0.0
Spleen 380	30.16	19.77	30.2	19.8	10.4	10.4	0.7
Kidney 27	32.55	21.02	32.6	21.0	11.5	11.5	0.3
Liver NDR 379	37.86	22.64	37.9	22.6	15.2	15.2	0.0
Fetal Liver BWH 54	35.04	23.3	35.0	23.3	11.7	11.7	0.3
Brain MCL 400	34.35	20.75	34.4	20.8	13.6	13.6	0.1
Colon PIT 259	38.03	22.89	38.0	22.9	15.1	15.1	0.0
mBM MNC LP7	38.68	19.38	38.7	19.4	19.3	19.3	0.0
mBM CD34+ LP92	35.59	21.08	35.6	21.1	14.5	14.5	0.0
mPB CD34+ LP350	34.41	20.35	34.4	20.4	14.1	14.1	0.1
mPB CD34+ LF53	32.51	19.51	32.5	19.5	13.0	13.0	0.1
BM CD34+ LF89	35.96	20.88	36.0	20.9	15.1	15.1	0.0
BM CD34+ LF75	40	24.06	40.0	24.1	15.9	15.9	0.0
Cord Blood CD34+ MF1	36.37	21.18	36.4	21.2	15.2	15.2	0.0
Cord Blood CD34+ LF101	40	20.09	40.0	20.1	19.9	19.9	0.0
GPA Hi LF154	33.87	22.39	33.9	22.4	11.5	11.5	0.4
GPA Hi LF156	30.53	21.14	30.5	21.1	9.4	9.4	1.5
GPA Lo LF154	37.86	23.44	37.9	23.4	14.4	14.4	0.0
GPA Lo LF156	33.39	22.02	33.4	22.0	11.4	11.4	0.4
MF11 Stromal D32 post irrad	38.27	19.03	38.3	19.0	19.2	19.2	0.0
MF13 Stromal D2 post irrad	33.62	18.11	33.6	18.1	15.5	15.5	0.0

TABLE 39


23565 mRNA expression

		Avg	Avg			Rel exp
69039	Beta	23565	Beta	ΔCT	ΔΔCT	Rel exp

BM CD61+LP196	31.11	22.28	31.1	22.3	8.8	8.8	2.2
Platelets LP57	33.89	17.1	33.9	17.1	16.8	16.8	0.0
mBM CD14-/11b-/15+ LF120	38.31	20.66	38.3	20.7	17.7	17.7	0.0
BM CD14-/11b-/15+ LF54	35.53	20.71	35.5	20.7	14.8	14.8	0.0
BM CD14-/11b-/15+ LF128	34.01	19.9	34.0	19.9	14.1	14.1	0.1
BM CD14-/11b-/15+ LF145	33.97	20.15	34.0	20.2	13.8	13.8	0.1
mBMCD14-/11b+/15+ LF120	36.12	20.22	36.1	20.2	15.9	15.9	0.0
BM CD14-/11b+/15+ LF106	36.95	20.32	37.0	20.3	16.6	16.6	0.0
BM-1 CD15+ ench LP41	35.23	19.32	35.2	19.3	15.9	15.9	0.0
Eryth D0 LF143	34.53	20.35	34.5	20.4	14.2	14.2	0.1
Eryth 48 hr LF143	33.67	21.44	33.7	21.4	12.2	12.2	0.2
Eryth D6 LF143	36.71	24.18	36.7	24.2	12.5	12.5	0.2
Eryth D12 LF143	35.15	22.74	35.2	22.7	12.4	12.4	0.2
Eryth D0 LF139	34.69	21.85	34.7	21.9	12.8	12.8	0.1
Eryth 24 hr LF139	40	23.77	40.0	23.8	16.2	16.2	0.0
Eryth D6 LF139	40	21.93	40.0	21.9	18.1	18.1	0.0
Eryth D12 LF139	36.39	23.5	36.4	23.5	12.9	12.9	0.1
BFU Eryth D7 LP79	31.51	22.07	31.5	22.1	9.4	9.4	1.4
BFU Eryth D7 LP95	33.39	22.97	33.4	23.0	10.4	10.4	0.7
BFU Eryth D7 +3epo LP81	35.44	22.62	35.4	22.6	12.8	12.8	0.1
BFU Eryth D7 +3epo LP104	34.41	23.22	34.4	23.2	11.2	11.2	0.4
Mast Cell LP118	40	21.18	40.0	21.2	18.8	18.8	0.0

TABLE 40


23565 mRNA expression

		Avg	Avg			Rel exp
69039	Beta	55503	Beta	ΔCT	ΔΔCT	Rel exp

Meg D0 LF140	33.87	19.13	33.87	19.13	14.74	14.74	0.036544
Meg 48 hr LF140	33.27	20.41	33	20	13	13	0
Meg D6 LF140	31.13	20.49	31	20	11	11	1
Meg D0 LF166	34.46	19.43	34	19	15	15	0
Meg 24 hr LF166	34.3	19.56	34	20	15	15	0
Meg 48 hr LF166	34.21	20.08	34	20	14	14	0
Meg D6 LF166	32.69	19.79	33	20	13	13	0
Meg D10 LF166	29.81	19.65	30	20	10	10	1
Neut D0 LF141	34.94	20.64	35	21	14	14	0
Neut 48 hr LF141	33.5	20.58	34	21	13	13	0
Neut D6 LF141	33.08	20.34	33	20	13	13	0
Neut D12 LF141	40	18.75	40	19	21	21	0
Neut D0 LF144	37.66	21.89	38	22	16	16	0
Neut 48 hr LF144	35.37	20.48	35	20	15	15	0
Neut D6 LF144	35.67	20.55	36	21	15	15	0
NeutD12LF144	40	20.54	40	21	19	19	0
NTC	40	40	40	40	0	0
NTC	40	40	40	40	0	0

Human 13305

The present invention is based, at least in part, on the discovery of novel molecules, referred to herein as “13305” nucleic acid and polypeptide molecules, which have homologies to known serine/threonine kinases at their active sites and in regions relating to ATP binding. Thus, 13305 proteins are expected to play a role in or function in signalling pathways associated with cellular growth.

The nucleotide sequence of the isolated human 13305 cDNA (SEQ ID NO:88), which is approximately 5389 nucleotides in length including untranslated regions, contains a predicted methionine-initiated coding sequence of about 3630 nucleotides, not including the termination codon (nucleotides 6-3635 of SEQ ID NO:88; 1-3630 of SEQ ID NO:90). The coding sequence encodes a 1210 amino acid protein (SEQ ID NO:89).

An alignment of the protein kinase family domain of human 13305 with a consensus amino acid sequence derived from a hidden Markov model (HMM) from PFAM shows the consensus amino acid sequence (SEQ ID NOs: 93-94) aligns with amino acids 190 to 411 and 492 to 518 of SEQ ID NO:89.

A BLAST alignment of human 13305 with a consensus amino acid sequence derived from a ProDomain “protein kinase nuclear serine/threonine-protein homeodomain-interacting homeobox DNA-binding serine/threonine F20B6.8” (Release 1999.2; see also ProDomain Release 2000.1) shows amino acid residues 1 to 158 of the 158 amino acid consensus sequence (SEQ ID NO:95) aligns with the “protein kinase nuclear serine/threonine-protein homeodomain-interacting homeobox DNA-binding serine/threonine F20B6.8” domain of human 13305, amino acid residues 416 to 565 of SEQ ID NO:89.

A BLAST alignment of human 13305 with a consensus amino acid sequence derived from a ProDomain “protein kinase nuclear homeodomain-interacting homeobox DNA-binding serine/threonine serine/threonine-protein” (Release 1999.2; see also ProDomain Release 2000.1) shows amino acid residues 72 to 272 of the amino acid consensus sequence (SEQ ID NOs: 96-98) aligns with the “protein kinase nuclear homeodomain-interacting homeobox DNA-binding serine/threonine serine/threonine-protein” domain of human 13305, amino acid residues 714 to 848, 720 to 887 an 615 to 667 of SEQ ID NO:89. The BLAST algorithm identifies multiple local alignments between the consensus amino acid sequence and human 13305.

A BLAST alignment of human 13305 with a consensus amino acid sequence derived from a ProDomain “protein kinase nuclear homeodomain-interacting homeobox DNA-binding serine/threonine serine/threonine-protein” (Release 1999.2; see also ProDomain Release 2000.1) shows amino acid residues 3 to 190 of the 190 amino acid consensus sequence (SEQ ID NO:99) aligns with the “protein kinase nuclear homeodomain-interacting homeobox DNA-binding serine/threonine serine/threonine-protein” domain of human 13305, amino acid residues 1030 to 1210 of SEQ ID NO:89.

Human 13305 contains the following regions or other structural features (for general information regarding PFAM identifiers, PS prefix and PF prefix domain identification numbers, refer to Sonnhammer et al. (1997) Protein 28:405-420: a eukaryotic protein kinase domain (PFAM Accession Number PF00069) located at about amino acid residues 190 to 411 and 492 to 518 of SEQ ID NO:89; three transmembrane domains (predicted by MEMSAT, Jones et al. (1994) Biochemistry 33:3038-3049) at about amino acids 73 to 89, 363 to 387, and 1156 to 1173 of SEQ ID NO:89; ten N-glycosylation sites (Prosite PS00001) from about amino acids 57 to 60, 111 to 114, 133 to 136, 149 to 152, 262 to 265, 471 to 474, 566 to 569, 570 to 573, 1009 to 1012 and 1045 to 1048 of SEQ ID NO:89; one glycosaminoglycan attachment sites (Prosite PS00002) from about amino acids 170 to 173 of SEQ ID NO:89; three cAMP/cGMP-dependent protein kinase phosphorylation sites (Prosite PS00004) located at about amino acids 124 to 127, 209 to 212, and 505 to 508 of SEQ ID NO:89; twelve protein kinase C phosphorylation sites (Prosite PS00005) at about amino acids 20 to 22, 107 to 109, 163 to 165, 211 to 213, 422 to 424, 666 to 668, 843 to 845, 853 to 855, 907 to 909, 1008 to 1010, 1138 to 1140 and 1187 to 1189 of SEQ ID NO:89; fifteen casein kinase II phosphorylation sites (Prosite PS00006) located at about amino acids 29 to 32, 37 to 40, 87 to 90, 113 to 116, 169 to 172, 211 to 214, 396 to 399, 441 to 444, 474 to 477, 643 to 646, 856 to 859, 910 to 913, 938 to 941, 967 to 970, and 1057 to 1060 of SEQ ID NO:89; one tyrosine kinase phosphorylation site. (Prosite PS00007) from about amino acids 452 to 459 of SEQ ID NO:89; seventeen N-myristoylation sites (Prosite PS00008) from about amino acids 35-40, 54-59, 93-98, 154-159, 310-315, 366-371, 379-384, 419-424, 662-667, 787-792, 800-805, 963-968, 1005-1010, 1019-1024, 1036-1041, 1124-1129 and 1186-1191 of SEQ ID NO:89; one ATP protein kinases ATP-binding region signature (Prosite PS00107) from about amino acids 196-204 of SEQ ID NO:89; and one serine-threonine protein kinases active site signature (Prosite PS00108) from about amino acids 311-323 of SEQ ID NO:89.

A hydropathy plot of human 13305 was performed. Polypeptides of the invention include fragments which include: all or part of a hydrophobic sequence, e.g., the sequence from about amino acid 300 to 310, from about 361 to 391, and from about 585 to 605 of SEQ ID NO:89; all or part of a hydrophilic sequence, e.g., the sequence from about amino acid 20 to 60, from about 245 to 265, and from about 220 to 260 of SEQ ID NO:89; a sequence which includes a Cys, or a glycosylation site.

The present invention is based, at least in part, on the discovery of novel molecules, referred to herein as 13305 protein and nucleic acid molecules, which comprise a family of molecules having certain conserved structural and functional features.

One embodiment of the invention features 13305 nucleic acid molecules, preferably human 13305 molecules, e.g., 13305. The 13305 nucleic acid and protein molecules of the invention are described in further detail in the following subsections.

In another embodiment, the isolated proteins of the present invention, preferably 13305 proteins, are identified based on the presence of at least Ser/Thr kinase site and at least one ATP-binding region.

As used herein, the term “Ser/Thr kinase site” includes an amino acid sequence of about 200-400 amino acid residues in length, preferably 200-300 amino acid residues in length, and more preferably 250-300 amino acid residues in length, which is conserved in kinases which phosphorylate serine and threonine residues and found in the catalytic domain of Ser/Thr kinases. Preferably, the Ser/Thr kinase site includes the following amino acid consensus sequence X ₉-g-X-G-X₄—V—X₁₂—K—X-(_10-19)-E-X₆₆-h-X₈-h-r-D-X—K—X₂—N—X₁₇—K—X₂-D-f-g-X₂₁-p-X₁₃-w-X₃-g-X₅₅—R—X₁₄-h-X₃(SEQ ID NO:91) (where invariant residues are indicated by upper case letters and nearly invariant residues are indicated by lower case letters). The nearly invariant residues are usually found in most Ser/Thr kinase sites, but can be replaced by other amino acids which, preferably, have similar characteristics. For example, a nearly invariant hydrophobic amino acid in the above amino acid consensus sequence would most likely be replaced by another hydrophobic amino acid. Ser/Thr kinase domains are described in, for example, Levin D. E. et al. (1990) Proc. Natl. Acad. Sci. USA 87:8272-76, the contents of which are incorporated herein by reference.

As used herein, the term “ATP-binding region” includes an amino acid sequence of about 20-40, preferably 20-30, and more preferably 25-30 amino acid residues in length, present in enzymes which activate their substrates by phosphorylation, and involved in binding adenosine triphosphate (ATP). ATP-binding regions preferably include the following amino acid consensus sequence: G-X-G-X—X-G-X(15-23)-K (SEQ ID NO:92). ATP-binding regions are described in, for example, Samuel K. P. et al. (1987) FEBS Lei. 218(1): 81-86, the contents of which are incorporated herein by reference. Amino acid residues 196 to 204 of SEQ ID NO:89 comprise an ATP-binding region. Amino acid residues 311-323 of the 13305 protein (SEQ ID NO:89) comprise a Ser/Thr kinase domain.

Isolated proteins of the present invention, preferably 13305 proteins, have an amino acid sequence sufficiently homologous to the amino acid sequence of SEQ ID NO:89 or are encoded by a nucleotide sequence sufficiently homologous to SEQ ID NO:88 or SEQ ID NO:89. The 13305 nucleic acid encodes a polypeptide with similarities to previously characterized protein kinases. Thus the 13305 encoded polypeptide is expected to be a kinase and function in the phosphorylation of protein substrates. The 13305 nucleic acid also encodes a polypeptide with similarities to previously identified homeodomains. Thus the 13305 encoded polypeptide is expected to be a kinase and function in the phosphorylation of proteins involved in interactions with DNA. The homeodomain of 13305 proteins may also be substituted for the homeodomains of other proteins in known assays based on the “swapping” of such domains.

As used interchangeably herein a “13305 activity”, “biological activity of 13305” or “functional activity of 13305”, refers to an activity exerted by a 13305 protein, polypeptide or nucleic acid molecule on a 13305 responsive cell or a 13305 protein substrate as determined in vivo, or in vitro, according to standard techniques. The biological activity of 13305 is described herein.

Accordingly, another embodiment of the invention features isolated 13305 proteins and polypeptides having a 13305 activity. Preferred proteins are 13305 proteins having at least one Ser/Thr kinase and at least one ATP-binding region. Additional preferred proteins have at least one Ser/Thr kinase site, at least one ATP-binding region, and preferably a 13305 activity. Additional preferred proteins have at least one Ser/Thr kinase site, at least one ATP-binding region, and are, preferably, encoded by a nucleic acid molecule having a nucleotide sequence which hybridizes under stringent hybridization conditions to a nucleic acid molecule comprising the nucleotide sequence of SEQ ID NO:88 or SEQ ID NO:90.

A 13305 polypeptide can include at least one, two, preferably three “transmembrane domains” or regions homologous with a “transmembrane domain”. As used herein, the term “transmembrane domain” includes an amino acid sequence of about 10 to 40 amino acid residues in length and spans the plasma membrane. Transmembrane domains are rich in hydrophobic residues, e.g., at least 50%, 60%, 70%, 80%, 90%, 95% or more of the amino acids of a transmembrane domain are hydrophobic, e.g., leucines, isoleucines, tyrosines, or tryptophans. Transmembrane domains typically have alpha-helical structures and are described in, for example, Zagotta, W. N. et al., (1996) Annual Rev. Neurosci. 19:235-263, the contents of which are incorporated herein by reference.

In a preferred embodiment, a 13305 polypeptide or protein has at least one, two, preferably three “transmembrane domains” or regions which includes at least about 12 to 35 more preferably about 14 to 30 or 15 to 25 amino acid residues and has at least about 60%, 70% 80% 90% 95%, 99%, or 100% homology with a “transmembrane domain,” e.g., the transmembrane domains of human 13305 (e.g., residues 73-89, 363-387, and 1156-1173 of SEQ ID NO:89). The transmembrane domain of human 13305 can be visualized in a hydropathy plot as regions of about 15 to 25 amino acids where the hydropathy trace is mostly above the horizontal line.

To identify the presence of a “transmembrane” domain in a 13305 protein sequence, and make the determination that a polypeptide or protein of interest has a particular profile, the amino acid sequence of the protein can be analyzed by a transmembrane prediction method that predicts the secondary structure and topology of integral membrane proteins based on the recognition of topological models (MEMSAT, Jones et al., (1994) Biochemistry 33:3038-3049).

A 13305 polypeptide can include at least one, two, three, preferably four “non-transmembrane regions.” As used herein, the term “non-transmembrane region” includes an amino acid sequence not identified as a transmembrane domain. The non-transmembrane regions in 13305 are located at about amino acids 1-72, 90-362, 388-1155, and 1174-1210 of SEQ ID NO:89.

The non-transmembrane regions of 13305 include at least one, preferably two cytoplasmic regions. In one embodiment, a cytoplasmic region of a 13305 protein can include the C-terminus and can be a “C-terminal cytoplasmic domain,” also referred to herein as a “C-terminal cytoplasmic tail.” As used herein, a “C-terminal cytoplasmic domain” includes an amino acid sequence having a length of at least about 5, preferably about 5 to 40, more preferably about 10 to 37 amino acid residues and is located inside of a cell or within the cytoplasm of a cell. The N-terminal amino acid residue of a “C-terminal cytoplasmic domain” is adjacent to a C-terminal amino acid residue of a transmembrane domain in a 13305 protein. For example, a C-terminal cytoplasmic domain is located at about amino acid residues 1174 to 1210 of SEQ ID NO:89.

In a preferred embodiment, a 13305 polypeptide or protein has a C-terminal cytoplasmic domain or a region which includes at least about 5, preferably about 5 to 40, and more preferably about 10 to 37 amino acid residues and has at least about 60%, 70% 80% 90% 95%, 99%, or 100% homology with a C-terminal cytoplasmic domain,” e.g., the C-terminal cytoplasmic domain of human 13305.(e.g., residues 1174 to 1210 of SEQ ID NO:89).

In another embodiment, a 13305 protein includes at least one, cytoplasmic loop. As used herein, the term “loop” includes an amino acid sequence that resides outside of a phospholipid membrane, having a length of at least about 5, preferably about 100 to 300, more preferably about 100 to 273 amino acid residues, and has an amino acid sequence that connects two transmembrane domains within a protein or polypeptide. Accordingly, the N-terminal amino acid of a loop is adjacent to a C-terminal amino acid of a transmembrane domain in a 13305 molecule, and the C-terminal amino acid of a loop is adjacent to an N-terminal amino acid of a transmembrane domain in a 13305 molecule. As used herein, a “cytoplasmic loop” includes a loop located inside of a cell or within the cytoplasm of a cell. For example, a “cytoplasmic loop” can be found at about amino acid residues 90-362 of SEQ ID NO:89.

In a preferred embodiment, a 13305 polypeptide or protein has a cytoplasmic loop or a region which includes at least about 4, preferably about 5, preferably about 100 to 300, more preferably about 100 to 273 amino acid residues and has at least about 60%, 70% 80% 90% 95%, 99%, or 100% homology with a cytoplasmic loop,” e.g., a cytoplasmic loop of human 13305 (e.g., residues 90-362 of SEQ ID NO:89).

In another embodiment, a 13305 protein includes at least one non-cytoplasmic loop. As used herein, a “non-cytoplasmic loop” includes an amino acid sequence located outside of a cell or within an intracellular organelle. Non-cytoplasmic loops include extracellular domains (i.e., outside of the cell) and intracellular domains (i.e., within the cell). When referring to membrane-bound proteins found in intracellular organelles (e.g., mitochondria, endoplasmic reticulum, peroxisomes microsomes, vesicles, endosomes, and lysosomes), non-cytoplasmic loops include those domains of the protein that reside in the lumen of the organelle or the matrix or the intermembrane space. For example, a “non-cytoplasmic loop” can be found at about amino acid residues 388-1155 of SEQ ID NO:89.

In a preferred embodiment, a 13305 polypeptide or protein has at least one non-cytoplasmic loop or a region which includes at least about 5, preferably about 100 to 800, more preferably about 100 to 768 amino acid residues and has at least about 60%, 70% 80% 90% 95%, 99%, or 100% homology with a “non-cytoplasmic loop,” e.g., at least one non-cytoplasmic loop of human 13305 (e.g., residues 388-1155 of SEQ ID NO:89).

The non-transmembrane regions of 13305 include at least one, “N-terminal extracellular domain.” As used herein, an “N-terminal extracellular domain” includes an amino acid sequence having about 1 to 100, preferably about 1 to 80, more preferably about 1 to 75, or even more preferably about 1 to 72 amino acid residues in length and is located outside of a cell or outside the cytoplasm of a cell. The C-terminal amino acid residue of an “N-terminal extracellular domain” is adjacent to an N-terminal amino acid residue of a transmembrane domain in a 13305 protein. For example, an N-terminal extracellular domain is located at about amino acid residues 1 to 72 of SEQ ID NO:89.

In a preferred embodiment, a polypeptide or protein has an N-terminal extracellular domain or a region which includes at least about 1 to 100, preferably about 1 to 80, more preferably about 1 to 72 amino acid residues and has at least about 60%, 70% 80% 90% 95%, 99%, or 100% homology with an “N-terminal extracellular domain,” e.g., the N-terminal extracellular domain of human 13305 (e.g., residues 1 to 72 of SEQ ID NO:89).

A 13305 family member can include at least one protein kinase domain; and at least one, !two, three, four, five, six, preferably seven transmembrane and non-transmembrane domains. Furthermore, a 13305 family member can include at least one, two, three, four, five, six, seven, eight, nine, preferably ten N-glycosylation sites (PS00001); at least one glycosaminoglycan attachement site (PS00002); at least one, two, preferably three cAMP/cGMP-dependent protein kinase phosphorylation sites (Prosite PS00004); at least one, two, three, four, five, six, seven, eight, nine, ten, eleven, preferably twelve protein kinase C phosphorylation sites (PS00005); at least one, two, three, preferably four casein kinase II phosphorylation sites (PS00006); at least one tyrosine kinase phosphorylation site (PS00007); at least one, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen and preferably fifteen N-myristoylation sites (PS00008); at least one ATP protein kinases ATP-binding region signature (PS00107); and at least one serine-threonine protein kinases active site signature (PS00108).

As used herein, the term “kinase domain” includes an amino acid sequence of about 100 to 275 amino acid residues in length and having a bit score for the alignment of the sequence to the kinase domain (HMM) of at least 100. Preferably a kinase domain mediates intracellular signal transduction. Preferably, a kinase domain includes at least about 100 to 275 amino acids, more preferably about 150 to 275 amino acid residues, or about 200 to 275 amino acids and has a bit score for the alignment of the sequence to the kinase domain (HMM) of at least 100, 150, 200, 250-or greater. The kinase domain (amino acids 190-411 and 492-518 of SEQ ID NO:89) of human 13305 align with a consensus amino acid sequence (SEQ ID NO:93-94) derived from a hidden Markov model. The “protein kinase” domain (HMM) has been assigned the PFAM Accession Number PF00069 and corresponds to about amino acids 190-7411 and 492-518 of SEQ ID NO:89.

In a preferred embodiment, a 13305 polypeptide or protein has a “kinase domain” or a region which includes at least about 100 to 215 more preferably about 150 to 275 or 200 to 275 amino acid residues and has at least about 60%, 70% 80% 90% 95%, 99%, or 100% homology with a “kinase domain,” e.g., the kinase domain of human 13305 (e.g., residues 190-411 and 492-518 of SEQ ID NO:89).

To identify the presence of a “kinase” domain in a 13305 protein sequence, and make the determination that a polypeptide or protein of interest has a particular profile, the amino acid sequence of the protein can be searched against the Pfam database of HMMs (e.g., the Pfam database, release 2.1) using the default parameters. For example, the hmmsf program, which is available as part of the HMMER package of search programs, is a family specific default program for MILPAT0063 and a score of 15 is the default threshold score for determining a hit. Alternatively, the threshold score for determining a hit can be lowered (e.g., to 8 bits). A description of the Pfam database can be found in Sonhammer et al. (1997) Proteins 28:405-420 and a detailed description of HMMs can be found, for example, in Gribskov et al. (1990) Meth. Enzymol.183:146-159; Gribskov et al. (1987) Proc. Natl. Acad. Sci. USA 84:4355-4358; Krogh et al. (1994) J. Mol. Biol. 235:1501-1531; and Stultz et al. (1993) Protein Sci. 2:305-314, the contents of which are incorporated herein by reference. A search was performed against the HMM database resulting in the identification of a “kinase domain” domain in the amino acid sequence of human 13305 at about residues 190-411 and 492-518 of SEQ ID NO:89.

To identify the presence of a “kinase” domain in a 13305 protein sequence, and make the determination that a polypeptide or protein of interest has a particular profile, the amino acid sequence of the protein can be searched against a database of domains, e.g., the ProDom database (Corpet et al. (1999), Nucl. Acids Res. 27:263-267). The ProDom protein domain database consists of an automatic compilation of homologous domains. Current versions of ProDom are built using recursive PSI-BLAST searches (Altschul S F et al. (1997) Nucleic Acids Res. 25:3389-3402; Gouzy et al. (1999) Computers and Chemistry 23:333-340) of the SWISS-PROT 38 and TREMBL protein databases. The database automatically generates a consensus sequence for each domain. A BLAST search was. performed against the HMM database resulting in the identification of a “kinase” domain in the amino acid sequence of human 13305 at about residues 416-465 of SEQ ID NO:89. The kinase domain is homologous to ProDom family “protein kinase nuclear serine/threonine-protein homeodomain-interacting homeobox DNA-binding serine/threonine F20B6.8,” SEQ ID NO:95, (ProDomain Release 1999.2). The consensus sequence for SEQ ID NO:95 is 72% identical over amino acids 416-465 of SEQ ID NO:89. The kinase domain is also homologous to ProDom family “protein kinase nuclear homeodomain-interacting homeobox DNA-binding serine/threonine serine/threonine-protein,” SEQ ID NO:96-98, (ProDomain Release 1999.2). The consensus sequences for SEQ ID NOs: 96-98 are 67%, 25% and 31% identical over amino acids 714 to 848, 720 to 887 and 615 to 667 of SEQ ID NO:89 respectively. The consensus sequences for SEQ ID NO:99 is 51% identical over amino acids 1030 to 1210 of SEQ ID NO:89.

In one embodiment, the 13305 molecules modulate the activity of one or more proteins involved in cellular growth or differentiation, e.g., brain, thymus, prostate epithelium, and fetal liver growth or differentiation. In another embodiment, the 13305 molecules of the present invention are capable of modulating the phosphorylation state of a 13305 molecule or one or more proteins involved in cellular growth or differentiation.

Additionally, 13305 nucleic acids and proteins have homology to known homeoboxes and homeodomains, respectively. Thus 13305 proteins are expected to exhibit DNA binding activity, in addition to kinase activity, under appropriate conditions. Without being bound by theory, 13305 protein may play a role in cellular function by being directed to appropriate locations based on the presence of the homeodomain, followed by providing its kinase activity to phosphorylate particular polypeptides at such locations. Possible roles for 13305 protein include developmental regulation.

Since the 13305 nucleic acid was found by TaqMan analysis to be expressed in cells of the brain, thymus, prostate epithelium, and fetal liver, the encoded protein kinase is at least expected to catalyze cell type specific phosphorylation reactions in those cells.

Additionally, the 13305 encoded protein kinase has homology to a mouse kinase orthologue. Thus, without being bound by theory, the 13305 kinase may be a human analogue of the mouse kinase.

As used herein, the term “protein kinase” includes a protein or polypeptide which is capable of modulating its own phosphorylation state or the phosphorylation state of another protein or polypeptide. Protein kinases can have a specificity for (i.e., a specificity to phosphorylate) serine/threonine residues, tyrosine residues, or both serine/threonine and tyrosine residues, e.g., the dual specificity kinases. As referred to herein, protein kinases preferably include a catalytic domain of about 200-400 amino acid residues in length, preferably about 200-300 amino acid residues in length, or more preferably about 250-300 amino acid residues in length, which includes preferably 5-20, more preferably 5-15, or preferably 11 highly conserved motifs or subdomains separated by sequences of amino acids with reduced or minimal conservation. Specificity of a protein kinase for phosphorylation of either tyrosine or serine/threonine can be predicted by the sequence of two of the subdomains (VIb and VIII) in which different residues are conserved in each class (as described in, for example, Hanks et al. (1988) Science 241:42-52) the contents of which are incorporated herein by reference). These subdomains are also described in further detail herein. Preferably, the kinases of the invention are serine/threonine kinases.

Protein kinases play a role in signalling pathways associated with cellular growth. For example, protein kinases are involved in the regulation of signal transmission from cellular receptors, e.g., growth-factor receptors; entry of cells into mitosis; and the regulation of cytoskeleton function, e.g., actin bundling. Thus, the 13305 molecules of the present invention may be involved in: 1) the regulation of transmission of signals from cellular receptors, e.g., cardiac cell growth factor receptors; 2) the modulation of the entry of cells into mitosis; 3) the modulation of cellular differentiation; 4) the modulation of cell death; and 5) the regulation of cytoskeleton function, e.g., actin bundling.

Further, 13305 molecules have been found by TaqMan analysis to be highly expressed in human bone marrow erythrocytes (GPA+ cells) and the human erythroleukemia cell line, K562, and has significant expression in GPA (low), erythroid progenitor cells. During erythroid differentiation, the expression of 13305 is regulated and 13305 has highest expression in terminally differentiated erythrocytes, which is expected for a kinase that negatively regulates cell growth. Inhibition of some dual-specificity kinases has been shown to enhance erythroid cell differentiation. As such, the 13305 molecules of the invention may play role in the regulation of erythroid cell growth, differentiation or both. For example, and without being bound by theory, it is expected that inhibition of 13305 activity in human bone marrow progenitor cells may lead to enhanced erythroid cell differentiation.

Additionally, 13305 molecules have been found to be overexpressed in tumor cells. Specifically, TaqMan analysis can be used to compare the expression levels in lung tumor cell lines versus a normal control and in multiple tumor cells versus normal tissue. Also, 13305 has shown increased expression in the A549 tumor cell line in S-phase (t=3). Without being bound by theory, it is likely that 13305 may be mutated and rendered inactive in tumor cells. Increased cell proliferation seen in tumor cells may be result of inactivity of 13305. Further, 13305 molecules may serve as specific and novel identifiers of such tumor cells.

Further, inhibition or over stimulation of the activity of protein kinases involved in signalling pathways associated with cellular growth can lead to perturbed cellular growth, which can in turn lead to cellular growth related disorders. As used herein, a “cellular growth related disorder” includes a disorder, disease, or condition characterized by a deregulation, e.g., an upregulation or a downregulation, of cellular growth. Cellular growth deregulation may be due to a deregulation of cellular proliferation, cell cycle progression, cellular differentiation and/or cellular hypertrophy.

Aberrant expression and/or activity of 13305 molecules may mediate disorders associated with bone metabolism

The 13305 nucleic acid and protein of the invention can be used to treat and/or diagnose a variety of immune disorders.

Disorders which may be treated or diagnosed by methods described herein include, but are not limited to, disorders associated with an accumulation in the liver of fibrous tissue, such as that resulting from an imbalance between production and degradation of the extracellular matrix accompanied by the collapse and condensation of preexisting fibers.

Additionally, 13305 may play an important role in the etiology of certain viral diseases and in the regulation of metabolism.

The 13305 molecules provide novel diagnostic targets and therapeutic agents to control pain in a variety of disorders, diseases, or conditions which are characterized by a deregulated, e.g., upregulated or downregulated, pain response.

Expression and Tissue Distribution of 13305

TaqMan real-time quantitative RT-PCR was used to detect the presence of RNA transcript corresponding to human 13305 in several tissues. It was found that the corresponding orthologs of 13305 are expressed in a variety of tissues. The presence of RNA transcript corresponding to human 13305 in RNA prepared from tumor and normal tissues was detected.

Transcriptional profiling results show an increased expression of 13305 mRNA in the lung tumor cell line, H460, in comparison with a normal human bronchial epithelium (NHBE) control. They also show the differential expression of 13305 RNA, in comparison with a NHBE control, in various lung tumor cell lines.

The expression of 13305 relative to the progression of cells through the cell cycle shows increased expression of 13305 RNA in S-phase (t=3) of the cell cycle in A549 cells.

Reverse Transcriptase PCR (RT-PCR) was used to detect the presence of RNA transcript corresponding to human 13305 in RNA prepared from tumor and normal tissues. Relative expression levels of the 13305 was assessed in breast, lung, colon and brain cells using TaqMan PCR and increased expression was found in 6/6 lung tumors, 3/8 breast tumors, and 3/4 colon tumor metastases in comparison to normal tissue controls. There is ubiquitous relative expression levels of 13305 in various tissues with significant expression in human fetal liver, thymus, prostate epithelial and brain cells.

In a hematology panel, there was expression of 13305 in human bone marrow erythrocytes (GPA+ cells), erythroid cells and the human erythroleukemia cell line, K562.

In another hematology panel, there was expression of 13305 in human bone marrow GPA+ cells and significant expression in GPA (low), erythroid progenitor cells.

Expression profiling results using in situ hybridization techniques have shown that 13305 mRNA has been detected in human lung and colon tumors. Low to moderate positive expression of 13305 has been shown in 3/3 lung tumor samples in comparison with 1/1 in normal lung tissue samples. Also, 13305 has been shown to be highly expressed in 4/4 primary colon tumor samples, and 2/3 colon tumor metastases, but not normal colon tissue samples (0/2).

As seen by these results, 13305 molecules have been found to be overexpressed in some tumor cells, and is presumably present in a mutated state and thus inactive. As such, 13305 molecules may serve as specific and novel identifiers of such tumor cells. Further, inhibitors of the 13305 molecules are also useful for the treatment of cancer, preferably lung cancer, and useful as a diagnostic.

Human 14911

The present invention is based, at least in part, on the discovery of novel molecules, referred to herein as “14911” nucleic acid and polypeptide molecules, which play a role in or function in the transduction of signals for cell proliferation, differentiation and apoptosis.

The human 14911 sequence (SEQ ID NO:100), which is approximately 1281 nucleotides in length, contains a predicted methionine-initiated coding sequence of about 1188 nucleotides, not including the termination codon (nucleotides 49-1236 of SEQ ID NO:100; 1-1188 of SEQ ID NO:102. The coding sequence encodes a 396 amino acid protein (SEQ ID NO:101).

A plasmid containing the nucleotide sequence encoding human 14911 was deposited with American Type Culture Collection (ATCC), 10801 University Boulevard, Manassas, Va. 20110-2209, on Jun. 7, 2001 and assigned Accession Number PTA-3435. This deposit will be maintained under the terms of the Budapest Treaty on the International Recognition of the Deposit of Microorganisms for the Purposes of Patent Procedure. This deposit was made merely as a convenience for those of skill in the art and is not an admission that a deposit is required under 35 U.S.C. §112.

A hydropathy plot of human 14911 shows relative hydrophobic residues and relative hydrophilic residues. The cysteine residues (cys) and N-glycosylation sites (Ngly) are alsoindicated.

The prediction of protein subcellular localization sites using PSORT software predicts the protein to be nuclear, followed by cytoplasmic or mitochondrial.

Results from the Prosite database of protein families and domains identify biologically significant sites. Human 14911 contains the following regions or other structural features: two N-glycosylation sites (PS00001) located at about amino acid residues 4 to 7 and 43 to 46 of SEQ ID NO:101; five protein kinase C phosphorylation sites (PS00005) located at about amino acid residues 5 to 7, 45 to 47, 122 to 124, 193 to 195 and 230 to 232 of SEQ ID NO:101; three casein kinase II phosproylation sites (PS00006) located at about amino acid residues 89 to 92, 212 to 215 and 230 to 233; three N-myristoylation sties (PS00008) located at about amino acid residues 2 to 7, 197 to 202 and 391 to 396 of SEQ ID NO:101; one amidation site (PS00009)located at about amino acid residues 218 to 221 of SEQ ID NO:101; one protein kinases ATP-binding region signature (PS00107) located at about amino acid residues 29 to 37 of SEQ ID NO:101; and one serine/threonine protein kinases active site signature (PS00108) located at about amino acid residues 142 to 154 of SEQ ID NO:101.

A PFAM search finds an “eukaryotic protein kinase domain” and a “protein kinase C-terminal domain” within human 14911. Amino acid residues 1 to 278 of the “eukaryotic protein kinase domain” consensus amino acid sequence (SEQ ID NO:105) align with amino acid residues 23 to 281 of SEQ ID NO:101. Amino acid residues 1 to 20 of the “protein kinase C terminal domain” consensus amino acid sequence (SEQ ID NO:106) align with amino acid residues 282 to 301 of SEQ ID NO:101.

Finally, a search of the ProDom protein domain database identifies homologous domains. The “kinase protein transferase ATP-binding serine/threonine-protein phosphorylation receptor tyrosine-protein precursor transmembrane” consensus amino acid sequence (SEQ ID NOs: 107-110) align with amino acid residues 23 to 71, 126 to 159, 172 to 312 and 250 to 280 of SEQ ID NO:101 over four HSPs. The “M03C11.1 protein” consensus amino acid sequence (SEQ ID NO:111) aligns with amino acid residues 280 to 372 of SEQ ID NO:101. The “F8K4.6 protein” consensus amino acid sequence (SEQ ID NO:112) aligns with amino acid residues 244 to 318 of SEQ ID NO:101.

In one embodiment, the 14911 molecules modulate the activity of one or more proteins involved in cellular growth or differentiation, e.g., cell growth or differentiation. In another embodiment, the 14911 molecules of the present invention are capable of modulating the phosphorylation state of a 14911 molecule or one or more proteins involved in cellular growth or differentiation.

In another embodiment, the isolated proteins of the present invention, preferably 14911 proteins, are identified based on the presence of at least one Ser/Thr kinase site and at least one ATP binding region.

As used herein, the term “Ser/Thr kinase site” includes an amino acid sequence of about 200-400 amino acid residues in length, preferably 200-300 amino acid residues in length, and more preferably 250-300 amino acid residues in length, which is conserved in kinases which phosphorylate serine and threonine residues and found in the catalytic domain of Ser/Thr kinases. Preferably, the Ser/Thr kinase site includes the following amino acid consensus sequence X ₉-g-X-G-X₄—V—X₁₂—K—X-(_10-19)-E-X₆₆-h-X₈-h-r-D-X—K—X₂—N—X₁₇—K—X₂-D-f-g-X₂₁-p-X₁₃-w-X₃-g-X₅₅—R—X₁₄-h-X₃(SEQ ID NO:103) (where invariant residues are indicated by upper case letters and nearly invariant residues are indicated by lower case letters). The nearly invariant residues are usually found in most Ser/Thr kinase sites, but can be replaced by other amino acids which, preferably, have similar characteristics. For example, a nearly invariant hydrophobic amino acid in the above amino acid consensus sequence would most likely be replaced by another hydrophobic amino acid. Ser/Thr kinase domains are described in, for example, Levin D. E. et al. (1990) Proc. Natl. Acad. Sci. USA 87:8272-76, the contents of which are incorporated herein by reference.

As used herein, the term “ATP-binding region” includes an amino acid sequence of about 20-40, preferably 20-30, and more preferably 25-30 amino acid residues in length, present in enzymes which activate their substrates by phosphorylation, and involved in binding adenosine triphosphate (ATP). ATP-binding regions preferably include the following amino acid consensus sequence: G-X-G-X—X-G-X(15-23)-K (SEQ ID NO:104). ATP-binding regions are described in, for example, Samuel K. P. et al. (1987) FEBS Let. 218(1): 81-86, the contents of which are incorporated herein by reference. Amino acid residues 31 to 39 of SEQ ID NO:101 comprise an ATP-binding region. Amino acid residues 144-156 of the 14911 protein (SEQ ID NO:101)comprise a Ser/Thr kinase domain.

The nucleic acid encodes a polypeptide with similarities known Ser/Thr kinases. Thus the 14911 encoded polypeptide is expected to be a kinase and function in the phosphorylation of protein substrates. Additionally, the 14911 nucleic acids can be used in known or novel screens and assays for kinase encoding nucleic acids to distinguish it from other distinct nucleic acids. Alternatively, the nucleic acid sequences can be used in the preparation of phylogenetic trees and relationships between organisms.

As used interchangeably herein a “14911 activity”, “biological activity of 14911” or “functional activity of 1491”, refers to an activity exerted by a 14911 protein, polypeptide or nucleic acid molecule on a 14911 responsive cell or a 14911 protein substrate as determined in vivo, or in vitro, according to standard techniques. The biological activity of 14911 is described herein.

Accordingly, another embodiment of the invention features isolated 14911 proteins and polypeptides having a 14911 activity. Preferred proteins are 14911 proteins having at least one Ser/Thr kinase and at least one ATP-binding region. Additional preferred proteins have at least one Ser/Thr kinase site, at least one ATP-binding region, and preferably a 14911 activity. Additional preferred proteins have at least one Ser/Thr kinase site, at least one ATP-binding region, and are, preferably, encoded by a nucleic acid molecule having a nucleotide sequence which hybridizes under stringent hybridization conditions to a nucleic acid molecule comprising the nucleotide sequence of SEQ ID NO:100 or SEQ ID NO:102.

As used herein, the term “protein kinase” includes a protein or polypeptide which is capable of modulating its own phosphorylation state or the phosphorylation state of another protein or polypeptide. Protein kinases can have a specificity for (i.e., a specificity to phosphorylate) serine/threonine residues, tyrosine residues, or both serine/threonine and tyrosine residues, e.g., the dual specificity kinases. As referred to herein, protein kinases preferably include a catalytic domain of about 200-400 amino acid residues in length, preferably about 200-300 amino acid residues in length, or more preferably about 250-300 amino acid residues in length, which includes preferably 5-20, more preferably 5-15, or preferably 11 highly conserved motifs or subdomains separated by sequences of amino acids with reduced or minimal conservation. Specificity of a protein kinase for phosphorylation of either tyrosine or serine/threonine can be predicted by the sequence of two of the subdomains (VIb and VIII) in which different residues are conserved in each class (as described in, for example, Hanks et al. (1988) Science 241:42-52) the contents of which are incorporated herein by reference). These subdomains are also described in further detail herein.

Protein kinases play a role in signaling pathways associated with cellular growth. For example, protein kinases are involved in the regulation of signal transmission from cellular receptors, e.g., growth-factor receptors; entry of cells into mitosis; and the regulation of cytoskeleton function, e.g., actin bundling. Thus, the 14911 molecules of the present invention may be involved in: 1) the regulation of transmission of signals from cellular receptors, e.g., growth factor receptors; 2) the modulation of the entry of cells into mitosis; 3) the modulation of cellular differentiation; 4) the modulation of cell death; and 5) the regulation of cytoskeleton function.

Additionally, and without being bound by theory, 14911 molecules have been found by TaqMan analysis to be overexpressed in tumor cells, where the molecules may be inappropriately propagating either cell proliferation or cell survival signals. As such, 14911 molecules may serve as specific and novel identifiers of such tumor cells. Further, inhibitors of the 14911 molecules are also useful for the treatment of cancer, preferably lung cancer, and useful as a diagnostic.

Inhibition or over stimulation of the activity of protein kinases involved in signaling pathways associated with cellular growth can lead to perturbed cellular growth, which can in turn lead to cellular growth related disorders. As used herein, a “cellular growth related disorder” includes a disorder, disease, or condition characterized by a deregulation, e.g., an upregulation or a downregulation, of cellular growth. Cellular growth deregulation may be due to a deregulation of cellular proliferation, cell cycle progression, cellular differentiation and/or cellular hypertrophy.

The present invention is based, at least in part, on the discovery of novel molecules, referred to herein as 14911 protein and nucleic acid molecules, which comprise a family of molecules having certain conserved structural and functional features.

One embodiment of the invention features 14911 nucleic acid molecules, preferably human 14911 molecules, e.g., 14911. The 14911 nucleic acid and protein molecules of the invention are described in further detail in the following subsections.

Expression and Tissue Distribution of 14911

TaqMan real-time quantitative RT-PCR was used to detect the presence of RNA transcript corresponding to human 14911 in several tissues. It was found that the corresponding orthologs of 14911 are expressed in a variety of tissues.

Relative expression levels of the 14911 was assessed in brain and lung cells using TaqMan PCR and increased expression was found in 2/5 lung tumor cell lines in comparison to a normal human bronchial epithelium (NHBE) control; 5/8 lung tumor samples in comparison to normal lung tissues; and 3/3 glioma samples in comparison to normal brain tissues. The relative expression levels and tissue distribution of the 14911 RNA was also assessed in a panel of human tissues or cells, including but not limited to heart, brain, breast, ovary, pancreas, prostate, colon, kidney, liver, fetal liver, lung, spleen, tonsil, lymph node, epithelial, endothelial, skeletal, fibroblasts, skin, adipose, bone cells (e.g., osteoclasts and osteoblasts), among others.

Expression profiling results using in situ hybridization techniques have shown that 14911 mRNA has been detected in human colon, lung, brain and breast tumors. Positive expression of 14911 has been shown in 2/4 lung tumors in comparison with lack of expression, 0/2, in normal lung tissue samples. Further, 14911 has been shown to be expressed both in tumors and normal tissues, specifically in 1/4 colon tumors and 1/1 normal colon tissue samples; 1/2 breast tumors and 1/1 normal breast tissue samples; and 1/3 brain tumors and 2/2 normal brain tissue samples.

As seen by these results, 1491 1 molecules have been found to be overexpressed in some tumor cells, where the molecules may be inappropriately propagating either cell proliferation or cell survival signals. As such, 14911 molecules may serve as specific and novel identifiers of such tumor cells. Further, inhibitors of the 14911 molecules are also useful for the treatment of cancer, preferably lung cancer, and useful as a diagnostic.

Human 86216

The present invention is based, in part, on the discovery of a novel DEAD helicase family member, referred to herein as “86216”.

The human 86216 sequence (SEQ ID NO:113), which is approximately 3577 nucleotides long including untranslated regions, contains a predicted methionine-initiated coding sequence of about 2337 nucleotides, not including the termination codon (nucleotides 47-2383 of SEQ ID NO:113; 1-2337 of SEQ ID NO:115). The coding sequence encodes a 779 amino acid protein (SEQ ID NO:114).

Human 86216 contains the following regions or other structural features (for general information regarding PFAM identifiers, PS prefix and PF prefix domain identification numbers, refer to Sonnhammer et al. (1997) Protein 28:405-420: a DEAD domain (PFAM Accession Number PF00270) located at about amino acid residues 117 to 231 of SEQ ID NO:114; a helicase c domain (PFAM Accession Number PF00271) located at about amino acid residues 300 to 401 of SEQ ID NO:114; a helicase RNA ATP dependent splicing domain (ProDom No. PD117102) located at about amino acid residues 213 to 300 of SEQ ID NO:114; one DEAH-box subfamily ATP-dependent helicases signature (SEQ ID NO:120) located at about amino acids 168 to 177 of SEQ ID NO:114; one ATP/GTP binding site motif A (P-loop) located at about amino acids 76 to 83 of SEQ ID NO:114; two N-glycosylation sites (Prosite PS00001) located at about amino acids 163 to 166 and 346 to 349 of SEQ ID NO:114; three cAMP/cGMP-dependent protein kinase phosphorylation sites (Prosite PS00004) located at about amino acids 427 to 430, 721 to 724, and 775 to 778 of SEQ ID NO:114; eight protein kinase C phosphorylation sites (Prosite PS00005) located at about amino acids 80 to 82, and 140 to 142, 394 to 396, 402 to 404, 437 to 439, 676 to 678, 724 to 726, and 774 to 776 of SEQ ID NO:114; nine casein kinase II phosphorylation sites (Prosite PS00006) located at about amino acids 43 to 46, 140 to 143, 308 to 311, 386 to 389, 485 to 488, 663 to 666, 724 to 727, 752 to 755, and 774 to 777 of SEQ ID NO:114; two tyrosine kinase phosphorylation sites (Prosite PS00007) located at about amino acids 248 to 255, and 526 to 532 of SEQ ID NO:114; six N-myristoylation sites (Prosite PS00008) located at about amino acids 48 to 53, 76 to 81, 127 to 132, 320 to 325, 337 to 342, and 567 to 572 of SEQ ID NO:114.

A hydropathy plot of human 86216 was performed. Polypeptides of the invention include fragments which include: all or part of a hydrophobic sequence, e.g., the sequence from about amino acid 205 to 225, from about 425 to 435, and from about 505 to 515 of SEQ ID NO:114; all or part of a hydrophilic sequence, e.g., the sequence from about amino acid 190 to 200, from about 530 to 540, and from about 710 to 730 of SEQ ID NO:114; a sequence which includes a Cys, or a glycosylation site.

The 86216 protein contains a significant number of structural characteristics in common with members of the DEAD helicase family. For example 86216 polypeptides of the invention conatain at least one DEAD domain, and at least one helicase-c domain.

As used herein, the term “DEAD helicase” includes a protein or polypeptide which is capable of ATP-dependent nucleic acid unwinding in eukaryotic cells.

Members of a DEAD helicase family of proteins in addition to their ability to unwind nucleic acids, are also involved in RNA metabolism, nuclear transcription, pre mRNA splicing, ribosome biogenesis, nucleocytoplasmic transport, translation, RNA decay, and organellar gene expression. The members of the DEAD superfamily share a number of conserved sequence motifs with those of the helicases superfamily. One of these motifs is the D-E-A-D- box, which is a version of a motif of ATP-binding proteins. Another subfamily of the ATP-dependent helicases has a conserved histidine instead of aspartic acid, and is referred to as a D-E-A-H box.

In addition, many proteins that bind ATP or GTP (e.g., DEAD helicases) share a glycine-rich region, which typically forms a flexible loop between a beta-strand and an alpha-helix. This loop interacts with one of the phosphate groups of the nucleotide. This sequence motif is generally referred to as the ‘A’ consensus sequence or the ‘P-loop’.

An alignment of the 86216 protein with a human RNA helicase gene of the DEAH-box protein family (SwissProt accession number O43143, corresponding to O43143 in Genbank) demonstrates about 38.1% sequence identity between the two sequences (as calculated in matblas from the blosum62.iij matrix).

A 86216 polypeptide can include a “DEAD domain” or regions homologous with a “DEAD domain”. A 86216 polypeptide can further include a “helicase-c domain” or regions homologous with a “helicase-c,” and at least one DEAH-box subfamily ATP-dependent helicases signature region.

A CLUSTAL W alignment shows amino acids 1 to 780 of human 86216 (SEQ ID NO:114) aligns with a human RNA helicase gene of the DEAH-box protein family, corresponding to amino acid residues 51 to 795 of SwissProt accession number 043143 in Genbank (SEQ ID NO:119). CLUSTAL W (v 1.74; Thompson et al. (1994) Nuc. Acids Res. 22:4673-80) uses dynamically varied gap penalties for progressive sequence alignments.

As used herein, the term “DEAD domain” includes an amino acid sequence of at least about 50 amino acid residues in length and having a bit score for the alignment of the sequence to the DEAD domain (HMM) of at least 1. Preferably a DEAD domain mediates ATP-dependent unwinding of nucleic acid. Preferably, a DEAD domain includes at least about 50 to 200 amino acids, more preferably about 75 to 150 amino acid residues, or most preferably about 90 to 120 amino acids and has a bit score for the alignment of the sequence to the DEAD domain (HMM) of at least 1, more preferably 3, and most preferably 4 or greater. A characteristic of the DEAD domain is the D-E-A-D- box, which is a conserved region characteristic of ATP-dependent helicases.

The DEAD domain can include a DEAH-box subfamily ATP-dependent helicases signature: [GSAH]—X-[LIVMF](3)-D-E-[ALIV]—H—[NECR] (SEQ ID NO:120).

In addition to the DEAH box subfamily ATP-dependent helicases signature in the DEAD domain, proteins belonging to this family of helicases also have an ATP/GTP-binding motif ‘A’ (P-loop) signature: [AG]-X(4)-G-K—[ST] (SEQ ID NO:121).

In the above conserved signature sequences, and other motifs or signature sequences described herein, the standard IUPAC one-letter code for the amino acids is used. Each element in the pattern is separated by a dash (-); square brackets ([]) indicate the particular residues that are accepted at that position; x indicates that any residue is accepted at that position; and numbers in parentheses (( )) indicate the number of residues represented by the accompanying amino acid.

The DEAD domain (HMM) has been assigned the PFAM Accession Number PF00270. An alignment of the DEAD domain of human 86216 with a consensus amino acid sequence derived from a hidden Markov model (HMM) from PFAM (SEQ ID NO:116) is derived from the hidden Markov model from Pfam and has a has a bit score of about 4.3.

The consensus amino acid sequence (SEQ ID NO:116), corresponds to amino acids 117 to 231 of SEQ ID NO:114.

In a preferred embodiment, a 86216 polypeptide or protein has a “DEAD domain” or a region which includes at least about 50 to 200 amino acids, more preferably about 75 to 150 amino acid residues, or most preferably about 90 to 120 amino acid residues and has at least about 60%, 70% 80% 90% 95%, 99%, or 100% homology with a “DEAD domain,” e.g., the DEAD domain of human 86216 (e.g., residues 117 to 231 of SEQ ID NO:114).

A 86216 polypeptide can include a “helicase-c domain” or regions homologous with a “helicase-c domain”. As used herein, the term “helicase-c domain” includes an amino acid sequence of at least about 50 amino acid residues in length and having a bit score for the alignment of the sequence to the helicase-c domain (HMM) of at least 1. Preferably a helicase-c domain is not restricted to the DEAD/DEAH helicases, and may be found in a wide variety of helicases and helicase related proteins. Preferably, a helicase-c domain includes at least about 50 to 150 amino acids, more preferably about 75 to 125 amino acid residues, or most preferably about 90 to 110 amino acids and has a bit score for the alignment of the sequence to the helicase-c domain (HMM) of at least −15, more preferably −10, and most preferably −5 or greater.

The helicase-c domain (HMM) has been-assigned the PFAM Accession Number PF00271. An alignment of the helicase-c domain (amino acids 300 to 401 of SEQ ID NO:114) of human 86216 with the Pfam helicase-c consensus amino acid sequence (SEQ ID NO:117) derived from a hidden Markov model yields a bit score of about −5.3.

In a preferred embodiment, a 86216 polypeptide or protein has a “helicase-c domain” or a region which includes at least about 50 to 150 amino acids, more preferably about 75 to 125 amino acid residues, or most preferably about 90 to 110 amino acid residues and has at least about 60%, 70% 80% 90% 95%, 99%, or 100% homology with a “helicase-c domain,” e.g., the helicase-c domain of human 86216 (e.g., residues 300 to 401 of SEQ ID NO:114).

To identify the presence of a “helicase-c” domain, and a “DEAD” domain in a 86216 protein sequence, and make the determination that a polypeptide or protein of interest has a particular profile, the amino acid sequence of the protein can be searched against the Pfam database of HMMs (e.g., the Pfam database, release 2.1) using the default parameters. For example, the hmmsf program, which is available as part of the HMMER package of search programs, is a family specific default program for MILPAT0063 and a score of 15 is the default threshold score for determining a hit. Alternatively, the threshold score for determining a hit can be lowered (e.g., to 8 bits). A description of the Pfam database can be found in Sonhammer et al. (1997) Proteins 28:405-420 and a detailed description of HMMs can be found, for example, in Gribskov et al. (1990) Meth. Enzymol: 183:146-159; Gribskov et al. (1987) Proc. Natl. Acad. Sci. USA 84:4355-4358; Krogh et al. (1994) J. Mol. Biol. 235:1501-1531; and Stultz et al. (1993) Protein Sci. 2:305-314, the contents of which are incorporated herein by reference. A search was performed against the HMM database resulting in the identification of a “helicase-c” domain in the amino acid sequence of human 86216 at about residues 300-401 of SEQ ID NO:114, and a “DEAD” domain in the sequence of human 86216 at about amino acid residues 117 to 231 of SEQ ID NO:114.

For further identification of domains, to identify the presence of a “helicase-c” domain in a 86216 protein sequence, and make the determination that a polypeptide or protein of interest has a particular profile, the amino acid sequence of the protein can be searched against a database of domains, e.g., the ProDom database (Corpet et al. (1999), Nucl. Acids Res. 27:263-267). The ProDom protein domain database consists of an automatic compilation of homologous domains. Current versions of ProDom are built using recursive PSI-BLAST searches (Altschul et al. (1997) Nucleic Acids Res. 25:3389-3402; Gouzy et al. (1999) Computers and Chemistry 23:333-340) of the SWISS-PROT 38 and TREMBL protein databases. The database automatically generates a consensus sequence for each domain.

A BLAST search was performed against the HMM database ProDomain Release 2001.1; resulting in the identification of a “Helicase RNA ATP-independent” No. PD 117102 domain (SEQ ID NO:118) in the amino acid sequence of human 86216 at about residues 213 to 301 of SEQ ID NO:114.

A 86216 family member can include at least one DEAD domain, and at least one helicase c domain. A 86216 family member can include at least one ATP/GTP binding site motif ‘A’ (P-loop) (Prosite PS00017), and at least one DEAH-box subfamily ATP-dependent helicase signature (Prosite PS00690).

Furthermore, a 86216 family member can include at least one, preferably two N-glycosylation site (Prosite PS00001); at least one, two, three, preferably four cAMP/cGMP protein kinase phosphorylation sites (Prosite PS00004); at least one, two, three, five, six, seven, preferably protein kinase C phosphorylation sites (Prosite PS00005); at least one, two, three, four, five, six, seven, eight, preferably nine casein kinase II phosphorylation sites (Prosite PS00006); at least one, preferably two tyrosine kinase phosphorylation sites (Prosite PS00007) and at least one, two, three, four, five, preferably six N-myristoylation sites (Prosite PS00008).

As the 86216 polypeptides of the invention can modulate 86216-mediated activities, they can be useful for developing novel diagnostic and therapeutic agents for DEAD helicase-associated or other 86216-associated disorders, as described below.

As used herein, a “DEAD helicase-associated activity” includes an activity which involves ATP-dependent, nucleic acid unwinding. Members of the DEAD helicase family can play a role in Bloom's syndrome, which is an autosomal recessive disorder associated with a predisposition to cancers of many types. Cells from those afflicted with Bloom's syndrome display extreme genomic instability. Helicase also may be implicated in Werner's syndrome (WS) another rare autosomal recessive disorder characterized by premature aging. Helicases may also have a role in breast cancer, wherein a member of the DEAH helicase family bearing a mutation in a residue (a residue known to be essential for catalytic function in other helicases), interferes with normal double-strand break repair.

As used herein, a “86216 activity”, “biological activity of 86216” or “functional activity of 86216”, refers to an activity exerted by a 86216 protein, polypeptide or nucleic acid molecule on e.g., a 86216-responsive cell or on a 86216 substrate, e.g., a protein substrate, as determined in vivo or in vitro. In one embodiment, a 86216 activity is a direct activity, such as an association with a 86216 target molecule. A “target molecule” or “binding partner” is a molecule with which a 86216 protein binds or interacts in nature. In an exemplary embodiment, 86216 is a helicase, e.g., a BACH1 helicase-like protein, which interacts directly with BRCA1 and contributes to its DNA repair (Cantor, SB et. al., (2001) Cell Apr 6;105(1):149-60) and thus binds to or interacts in nature with a molecule(or protein substrate), e.g., a nucleic acid binding protein.

A 86216 activity can also be an indirect activity, e.g., a cellular signaling activity mediated by interaction of the 86216 protein with a 86216 receptor. Based on the above-described sequence structures and similarities to molecules of known function, the 86216 molecules of the present invention can have similar biological activities as DEAD helicase family members. For example, the 86216 proteins of the present invention can have the ability to modulate any one or more of the following activities: (1) ATP dependent nucleic acid unwinding (2) the ability to modulate cellular proliferative disorders (e.g., proliferative disorders of the breast (e.g., breast cancer (e.g., proliferative disoprders of mammary epithelial cells) Bloom's syndrome, or Werners syndrome)); (3) RNA metabolism (e.g., nuclear transcription, and mRNA splicing); (4) nucleocytoplasmic transport; and (5) RNA decay and organellar expression.

The 86216 molecules of the invention can modulate the activities of cells in tissues where they are expressed. For example, TaqMan analysis shows 86216 mRNA is expressed in the mammary epithelial cell line MCF10A. Thus, the 86216 molecules can be used to treat breast disorders (i.e., proliferative cell disorders of the breast) in part because the 86216 mRNA is expressed in breast derived cells.

Thus, the 86216 molecules can act as novel diagnostic targets and therapeutic agents for controlling one or more disorders of the breast or other DEAD helicase disorders. As used herein, “DEAD helicase disorders” are diseases or disorders whose pathogenesis is caused by, is related to, or is associated with aberrant or deficient DEAD helicase protein function or expression. Examples of such disorders, e.g., DEAD helicase-associated or other 86216-associated disorders, include but are not limited to breast disorders and cellular proliferative and/or differentiative disorders.

The 86216 molecules can be used to treat breast disorders in part because DEAD helicase family members are found in mammary epithelial cells.

The 86216 molecules of the invention can be used to monitor, treat and/or diagnose a variety of proliferative disorders. Such disorders include hematopoietic neoplastic disorders.

Gene Expression Analysis of 86216

Human 86216 expression was measured by TaqMan® quantitative PCR (Perkin Elmer Applied Biosystems) in cDNA prepared from a variety of normal and diseased (e.g., cancerous) human tissues or cell lines.

The results indicate significant 86216.expression in the mammary epithelial cell line MCF10a.

Human 25206

The present invention is based, in part, on the discovery of a novel short-chain dehydrogenase/reductase, referred to herein as “25206”.

The human 25206 sequence (SEQ ID NO:122), which is approximately 1649 nucleotides long including untranslated regions, contains a predicted methionine-initiated coding sequence of about 858 nucleotides (nucleotides 213-1070 of SEQ ID NO:122; 1-858 of SEQ ID NO:124), not including the termination codon. The coding sequence encodes a 286 amino acid protein (SEQ ID NO:123).

The human 25206 protein of SEQ ID NO:123 includes an amino-terminal hydrophobic amino acid sequence, consistent with a signal sequence, of about 19 amino acids (from amino acid 1 to about amino acid 19 of SEQ ID NO:123), which upon cleavage results in the production of a mature protein form of 267 amino acids (from about amino acid 20 to about amino acid 286 of SEQ ID NO:123).

An alignment of the short-chain dehydrogenase/reductase domain of human 25206 with a consensus amino acid sequence derived from a hidden Markov model (HMM) from PFAM shows the consensus amino acid sequence (SEQ ID NO:125) aligns with amino acids 30 to 216 of SEQ ID NO:123.

Human 25206 contains the following regions or other structural features: a short-chain dehydrogenase/reductase domain (PFAM Accession Number PF00106) located at about amino acid residues 30 to 216 of SEQ ID NO:123, which includes a short-chain alcohol dehydrogenase family signature (PS00061) located at about amino acid residues 178 to 188 of SEQ ID NO:123; a signal peptide from about amino acids 1-19 of SEQ ID NO:123; two predicted Protein Kinase C phosphorylation sites (PS00005) at about amino acids 146 to 148 and 191 to 193 of SEQ ID NO:123; two predicted Casein Kinase II phosphorylation sites (PS00006) located at about amino acids 152 to 155 and 217 to 220 of SEQ ID NO:123; one predicted N-glycosylation site (PS00001) from about amino acids 280 to 283 of SEQ ID NO:123; and three predicted N-myristoylation sites (PS00008) from about amino acids 36 to 41, 117 to 122, and 244 to 249 of SEQ ID NO:123.

A hydropathy plot of human 25206 was performed. Polypeptides of the invention include fragments which include: all or part of a hydrophobic sequence, e.g., the sequence from about amino acid 76 to 88, from about 155 to 170, and from about 198 to 211 of SEQ ID NO:123; all or part of a hydrophilic sequence, e.g., the sequence of from about amino acid 120 to 131, from about 190 to 197, and from about 265 to 279 of SEQ ID NO:123.

The 25206 protein contains a significant number of structural characteristics in common with members of the short-chain dehydrogenase/reductase family.

Dehydrogenases typically contain at least two domains, the first binds a coenzyme, such as NAD or NADP, and the second binds substrate. Sequence of the coenzyme domain does not appear to be conserved among dehydrogenases. The second domain determines substrate specificity and contains amino acids involved in catalysis. Members of this family include alchohol dehydrognase, 3-β-hydroxysteroid dehydrogenase, estradiol 17-β-dehydrogenase, retinal dehydrogenase, and NADPH-dependent carbonyl reductase.

Short-chain dehydrogenases/reductases (SDRs) typically function as dimers or tetramers. The subunits are composed of approximately 250 to 300 amino acid residues, an N-terminal co-enzyme binding pattern of GxxxGxG (SEQ ID NO:126), and an active-site pattern of YxxK (SEQ ID NO:127) (Opperman et al. (1999) Enzymology and Molecular Biology of Carbonyl Metabolism 7 ed. Weiner et al., Plenum Publishers, NY p. 373-377). Although identity between different SDR members is at the 15-30% level, three-dimensional structures thus far analyzed reveal a highly similar conformation with a one-domain subunit with seven to eight β-strands.

25206 polypeptides are homologous to 11-beta hydroxysteroid dehydrogenase (11 beta-HSD), alternatively known as corticosteroid 11-beta dehydrogenase. Two isoforms of 11-beta HSD are known (Krozowski, Z. et al. (1999) J. Steroid Biochem. Mol. Biol. 69(1-6):391-401). These enzymes catalyze the interconversion of cortisol and the inactive glucocorticoid metabolite cortisone in an NADPH-dependent manner. 25206 polypeptide is closely related to the type I isoform, which is a bi-directional enzyme acting predominantly as a reductase to convert inactive cortisone to active cortisol. The type II isoform acts unidirectionally to inactivate cortisol.

A 25206 polypeptide can include a “short chain dehydrogenase domain” or regions homologous with a “short chain dehydrogenase domain”. Short chain dehydrogenases have the ability to directly or indirectly remove a hydride from a substrate, e.g., an alcohol; an aldehyde; a steroid, e.g., a glucocorticoid, cortisone; a sugar. Typically, after removal of a hydride from a substrate, electrons of the hydride are transferred to NAD+, NADP+, or other coenzyme (e.g., 3-acetylpyridine adenine dinucleotide phosphate) or hydride acceptor. For example, if the substrate has hydroxyl, dehydrogenation converts the hydroxyl to a keto group and produces NADH or NADPH and a proton. Hydride removal from substrate however does not require the presence of an acceptor. Free hydride can be detected, for example, optically by H+ binding to a dye molecule.

A 25206 polypeptide can include a “short-chain dehydrogenase/reductase domain” or regions homologous with a “short-chain dehydrogenase/reductase domain”.

As used herein, the term “short chain dehydrogenase domain” includes an amino acid sequence of about 50 to 400 amino acid residues in length and having a bit score for the alignment of the sequence to the short chain dehydrogenase domain (HMM) of at least 50. Preferably, a short chain dehydrogenase domain includes at least about 100 to 300 amino acids, more preferably about 140 to 250 amino acid residues, or about 180 to 190 amino acids and has a bit score for the alignment of the sequence to the short chain dehydrogenase domain (HMM) of at least 80, 100, 110 or greater. The short chain dehydrogenase domain (HMM) has been assigned the PFAM Accession Number PF00106. The short chain dehydrogenase domain (amino acids 30 to 216 of SEQ ID NO:123) of human 25206 aligns with a consensus amino acid sequence (SEQ ID NO:125) derived from a hidden Markov model.

In a preferred embodiment, 25206 polypeptide or protein has a “short chain dehydrogenase domain” or a region, that includes at least about 100 to 300 amino acids, more preferably about 140 to 250 amino acid residues, or about 180 to 190 amino acid residues and has at least about 60%, 70% 80% 90% 95%, 99%, or 100% homology with a “short chain dehydrogenase domain,” e.g., the short chain dehydrogenase domain of human 25206 (e.g., residues 30 to 216 of SEQ ID NO:123).

Preferably, the short chain dehydrogenase domain of a 25206 polypeptide includes a short chain dehydrogenase family signature, YSAAKF, ALDGF (SEQ ID NO:128), which corresponds to amino acids 178-188 of SEQ ID NO:123.

To identify the presence of a “short-chain dehydrogenase/reductase” domain in a 25206 protein sequence, and make the determination that a polypeptide or protein of interest has a particular profile, the amino acid sequence of the protein can be searched against the Pfam database of HMMs (e.g., the Pfam database, release 2.1) using the default parameters. For example, the hmmsf program, which is available as part of the HMMER package of search programs, is a family specific default program for MWLPAT0063 and a score of 15 is the default threshold score for determining a hit. Alternatively, the threshold score for determining a hit can be lowered (e.g., to 8 bits). A description of the Pfam database can be found in Sonhammer et al. (1997) Proteins 28(3):405-420 and a detailed description of HMMs can be found, for example, in Gribskov et al.(1990) Meth. Enzymol. 183:146-159; Gribskov et al.(1987) Proc. Natl. Acad. Sci. USA 84:4355-4358; Krogh et al.(1994) J. Mol. Biol. 235:1501-1531; and Stultz et al.(1993) Protein Sci. 2:305-314, the contents of which are incorporated herein by reference. A search was performed against the HMM database resulting in the identification of a “short-chain dehydrogenase/reductase” domain in the amino acid sequence of human 25206 at about residues 30 to 216 of SEQ ID NO:123.

A 25206 family member can include one or more of: a short chain dehydrogenase domain or a short chain alcohol dehydrogenase family signature. Furthermore, a 25206 family member can include a signal peptide; at least one, and preferably two, protein kinase C phosphorylation sites (PS00005); at least one, and preferably two, predicted casein kinase II phosphorylation sites (PS00006); and at least one predicted N-myristoylation sites (PS00008).

In yet another embodiment, the 25206 molecule can further include a signal sequence. As used herein, a “signal sequence” refers to a peptide of about 10-40 amino acid residues in length which occurs at the N-terminus of secretory and integral membrane proteins and which contains a majority of hydrophobic amino acid residues. For example, a signal sequence contains at least about 15-30 amino acid residues, preferably about 19 amino acid residues, and has at least about 40-70%, preferably about 50-65%, and more preferably about 55-60% hydrophobic amino acid residues (e.g., alanine, valine, leucine, isoleucine, phenylalanine, tyrosine, tryptophan, or proline). Such a “signal sequence”, also referred to in the art as a “signal peptide”, serves to direct a protein containing such a sequence to a lipid bilayer. For example, in one embodiment, a 25206 protein contains a signal sequence of about amino acids 1-19 of SEQ ID NO:123. The “signal sequence” is cleaved during processing of the mature protein. The mature 25206 protein corresponds to amino acids 20 to 286 of SEQ ID NO:123.

As the 25206 polypeptides of the invention may modulate 25206-mediated activities, they may be useful for developing novel diagnostic and therapeutic agents for 25206-mediated or related disorders, as described below.

As used herein, a “25206 activity”, “biological activity of 25206” or “functional activity of 25206”, refers to an activity exerted by a 25206 protein, polypeptide or nucleic acid molecule. For example, a 25206 activity can be an activity exerted by 25206 in a physiological milieu on, e.g., a 25206-responsive cell or on a 25206 substrate, e.g., a protein substrate. A 25206 activity can be determined in vivo or in vitro. In one embodiment, a 25206 activity can be an indirect activity, e.g., a cellular signaling activity mediated by interaction of the 25206 protein with a 25206 receptor.

In other embodiments, the 25206 activity is a direct activity, such as an association with a 25206 target molecule. A “target molecule” or “binding partner” is a molecule with which a 25206 protein binds or interacts in nature. For example, a 25206 binding partner is a substrate, e.g., an alcohol; an aldehyde; a steroid, e.g., a glucocorticoid, cortisone; a sugar. As the 25206 polypeptides show structural similarity to 11-beta-HSD, these polypeptides may be involved in the metabolism of steroids, e.g., glucocorticoids. Glucocorticoids have been shown to have an antiproliferative effect on some breast cancer cell lines in vitro (Hundertmark, S. et al. (1997) J. Endocrinol. 155(1):171-180). Accordingly, the 25206 molecules of the present invention may be involved in regulating cellular proliferation and differentiation.

Based on the above-described sequence similarities, the 25206 molecules of the present invention are predicted to have similar biological activities as short chain dehydrogenase family members. For example, the 25206 proteins of the present invention can have one or more of the following activities: (1) steroid biosynthesis or metabolism (breakdown); (2) changes associated with steroid biosynthesis or metabolism (e.g., sex trait development); (3) metabolism or removal of natural or xenobiotic substances (e.g., ethanol, toxins, etc.); (4) cellular proliferation or differentiation; or (5) cellular survival and/or degeneration (e.g., neurodegeneration).

As described below, TaqMan analysis shows 25206 mRNA is expressed in cancerous tissues, e.g., cancerous tissues from the breast, brain, lung, colon, liver, as well as neural (e.g., brain) or reproductive, e.g., ovarian, tissues. Thus, the 25206 molecules can act as novel diagnostic targets and therapeutic agents for controlling one or more of cellular proliferative, differentiative, neural, e.g., neurodegenerative, and reproductive, disorders.

Examples of cellular proliferative and/or differentiative disorders include cancer, e.g., carcinoma, sarcoma, metastatic disorders or hematopoietic neoplastic disorders, e.g., leukemias. A metastatic tumor can arise from a multitude of primary tumor types, including but not limited to those of prostate, colon, lung, brain, breast and liver origin.

Additional examples of proliferative disorders include hematopoietic neoplastic disorders.

Tissue Distribution of 25206 mRNA by TaqMan Analysis

Endogenous human 25206 gene expression was determined using the Perkin-Elmer/ABI 7700 Sequence Detection System which employs TaqMan technology.

Tissues tested include the human tissues and several cell lines shown in Tables 41-44. 25206 mRNA was detected in brain tissue (normal and tumorigenic), breast tissue (normal and tumorigenic), ovarian tissue (normal and tumorigenic), lung tissue (normal and tumorigenic), a host of xenograft cells and a host of breast cell clones (Tables 41-44). More specifically, as depicted in Tables 41-44, 25206 mRNA expression was increased 1.5-3.6 fold at all timepoints following IGF1 treatment. Additionally, 25206 mRNA was significantly upregulated in two MCF10AT3B tumor cell clones grown in soft agar vs. grown on plastic. 25206 mRNA was upregulated about 3 fold in 2/7 breast tumors vs. 3/4 normal breast tissues, and 3/7 lung tumors vs. 4/4 normal lung tissues. Phase I Taqman panel showed highest expression in brain tissue. 25206 showed expression in many tumor cell lines (NCIH67>A549>T47D). Each of these tables is described in more detail below.

Table 41 depicts the relative expression of 25206:mRNA in a panel of human tissues indicated below. Tissues depicted with as MET are metastatic tissue; HMVEC cells are human microvascular endothelial cells. 25206 mRNA is overexpressed in normal brain tissue and to some extent in tumorigenic brain (glioma) tissue.

TABLE 41


		Relative
Tissue source	Tissue Type	Expression

CHT 396	Colon Normal	0.0
CHT 519	Colon Normal	0.0
CHT 416	Colon Normal	0.1
CHT 452	Colon Normal	0.0
CHT 398	Colon Tumor	0.2
CHT 807	Colon Tumor	0.0
CHT 805	Colon Tumor	0.3
CHT 528	Colon Tumor	0.1
CHT 368	Colon Tumor	0.0
CHT 372	Colon Tumor	0.3
CHT 01	Liver Met	0.1
CHT 3	Liver Met	0.4
CHT 896	Liver Met	0.1
CHT 340	Liver Met	0.5
PIT 260	Liver Normal	0.0
PIT 229	Liver Normal	2.4
MGH 16	Brain Normal	29.8
MCL 53	Brain Normal	99.4
MCL 377	Brain Normal	26.8
MCL 390	Brain Normal	67.9
MPI 665	Astrocytes	6.7
CHT 201	Glio	0.6
CHT 216	Glio	5.7
CHT 501	Glio	5.6
CHI 1273	Glio	37.4
CHT 828	Glio	2.9
A24	HMVEC-Arr	1.1
C48	HMVEC-Prol	1.0
CHT 50	Placenta	0.4
BWH 58	Fetal Adrenal	8.9
PIT 251	Fetal Adrenal	1.7
BWH 54	Fetal Liver	0.7
BWH 75	Fetal Liver	0.4
	NTC	1000.0

Table 42 depicts the relative expression of 25206 mRNA in a panel of human tissues indicated below. 25206 mRNA is relatively overexpressed in breast, ovary, and lung tumorigenic tissue, while the gene is also overexpressed in normal ovary tissue.

	TABLE 42


		Relative
	Tissue	Expression

	Breast Normal	0.8
	Breast Normal	1.4
	Breast Normal	2.9
	Breast Normal	0.7
	Breast Tumor	2.1
	Breast Tumor	0.9
	Breast Tumor	0.3
	Breast Tumor	0.4
	Breast Tumor	1.8
	Breast Tumor	5.0
	Breast Tumor	4.1
	Ovary Normal	6.9
	Ovary Normal	6.1
	Ovary Normal	7.5
	Ovary Normal	7.9
	Ovary Tumor	0.6
	Ovary Tumor	0.6
	Ovary Tumor	6.1
	Ovary Tumor	1.5
	Ovary Tumor	2.2
	Ovary Tumor	0.2
	Ovary Tumor	7.3
	Ovary Tumor	0.4
	Lung Norm	0.3
	Lung Norm	0.9
	Lung Norm	0.3
	Lung Norm	0.5
	Lung Tumor	3.0
	Lung Tumor	3.1
	Lung Tumor	2.2
	Lung Tumor	1.4
	Lung Tumor	14.5
	Lung Tumor	1.7
	Lung Tumor	0.3

Table 43 depicts the relative expression of 25206 mRNA in a panel of human breast cell lines indicated below. Breast carcinoma cell lines are represented by MCF10, MCF-7, ZR, T47, MDA, and SKBr3. Normal breast cells are represented by the cell line Hs578. Expression of 25206 mRNA is upregulated in breast carcinoma cells grown in soft agar compared to breast carcinoma cells grown on plastic. Exposure of the MCF10 carcinoma line with insulin-like growth factor 1 (IGF-1) or epidermal growth factor (EGF) had some effect on the expression of 25206 mRNA.

	TABLE 43


	Tissue Type	Expression

	MCF10MS	1.09
	MCF10A	1.23
	MCF10AT.cl1	0.42
	MCF10AT.cl3	0.63
	MCF10AT1	0.49
	MCF10AT3B	0.73
	MCF10CA1a.cl1	0.41
	MCF10AT3B Agar	11.13
	MCF10ACA1a.cl1 Agar	2.03
	MCF10A.m25 Plastic	2.13
	MCF10CA Agar	1.52
	MCF10CA Plastic	0.46
	MCF3B Agar	6.24
	MCF3B Plastic	0.92
	MCF10A EGF 0 hr	0.50
	MCF10A EGF 0.5 hr	0.42
	MCF10A EGF 1 hr	0.37
	MCF10A EGF 2 hr	0.37
	MCF10A EGF 4 hr	0.43
	MCF10A EGF 8 hr	0.41
	MCF10A IGF1A 0 hr	0.76
	MCF10A IGF1A 0.5 hr	1.09
	MCF10A IGF1A 1 hr	1.02
	MCF10A IGF1A 3 hr	1.46
	MCF10A IGF1A 24 hr	2.74
	MCF10AT3B.cl5 Plastic	1.54
	MCF10AT3B.cl6 Plastic	0.95
	MCF10AT3B.cl3 Plastic	0.95
	MCF10AT3B.cl1 Plastic	0.88
	MCF10AT3B.cl4 Plastic	0.75
	MCF10AT3B.cl2 Plastic	0.67
	MCF10AT3B.cl5 Agar	9.49
	MCF10AT3B.cl6 Agar	10.49
	MCF-7	0.75
	ZR-75	1.58
	T47D	1.31
	MDA-231	0.35
	MDA-435	1.12
	SkBr3	0.13
	Hs578Bst	0.68
	Hs578T	0.62

Table 44 depicts the relative expression of 25206 mRNA in panel of human cancer cell lines after transplantation into mice. Human breast carcinoma cells lines are represented by MCF, ZR75, T47D, MDA, and SKBr3 cell lines; colon carcinoma cell lines are represented by DLD, SW620, HCT116 and Colo205 cell lines; lung adenosquamous carcinoma cell lines are represented by NCIH125, NCIH-67, NCIH 322, and NCIH460 cell lines; a lung carcinoma cell line is represented by A549 cell line; a lung cell line is represented by NHBE cell lines; ovarian carcinoma cells are represented by SKOV and OVCAR cell lines; and baby kidney cells which are indicated below. 25206 mRNA shows a slight increase in expression in all lung cell lines (both cancerous and normal), but is greatly overexpressed in baby kidney cells.

	TABLE 44


		Relative
	Tissue Type	Expression

	MCF-7 Breast T	4.69
	ZR75 Breast T	4.61
	T47D Breast T	6.87
	MDA 231 Breast T	2.21
	MDA 435 Breast T	6.64
	SKBr3 Breast	0.94
	DLD 1 Colon T (stageC)	2.98
	SW620 Colon T (stageC)	2.07
	HCT116	3.33
	HT29	0.22
	Colo 205	0.13
	NCIH125	3.93
	NCIH67	10.13
	NCIH322	7.16
	NCIH460	1.58
	A549	8.91
	NHBE	9.42
	SKOV-3 ovary	1.28
	OVCAR-3 ovary	4.74
	293 baby kidney	15.63
	293T baby kidney	24.77

Additional expression studies were conducted using probes generated from 4 normal breast tissue samples, 4 ductal carcinoma in situ (DCIS) samples, 4 invasive ductal carcinoma (IDC) samples and 3 invasive Iobular carcinoma (ILC) samples. 25206 mRNA was expressed at about 2 fold the median value of the 4 normal breast samples in 1/4 DCIS samples, 1/4 IDC samples and 0/3 ILC samples.

mRNA expression was assayed with probes generated from untreated human breast epithelial MCF10A cells or MCF10A cells treated with 10 nM IGF1 for 0.5, 1, 3 and 26 hours. 25206 mRNA expression was increased 1.5-1.8 fold at all timepoints following IGF1 treatment.

Tissue Distribution of 25206 mRNA by In Situ Hybridization

In situ hybridization studies revealed expression of 25206 mRNA in the following tissues: 0/2 normal breast tissues, 1/5 breast tumors, 0/3 normal lung tissues, 1/4 lung tumors, 0/1 normal colon tissue, 0/3 colon tumors, 0/1 normal ovary tissue, 0/2 ovary tumors and 1/1 normal brain tissue.

Human 8843

The present invention is based, in part, on the discovery of a novel dual specificity phosphatase family member, referred to herein as “8843”.

The human 8843 sequence (SEQ ID NO:129), which is approximately 839 nucleotides long including untranslated regions, contains a predicted methionine-initiated coding sequence of about 603 nucleotides, not including the termination codon (nucleotides 44-646 of SEQ ID NO:129; 1-603 of SEQ ID NO:131). The coding sequence encodes a 201 amino acid protein (SEQ ID NO:130).

An alignment of the dual specificity phosphatase domain of human 8843 with a consensus amino acid sequence derived from a hidden Markov model (HMM) from PFAM shows the consensus amino acid sequence (SEQ ID NO:132) aligns with amino acids 37 to 185 of SEQ ID NO:130.

An alignment of the dual specificity phosphatase domain of human 8843 with a consensus amino acid sequence derived from a hidden Markov model (HMM) in the SMART domain library shows the consensus amino acid sequence (SEQ ID NO:133) aligns with amino acids 37 to 185 of SEQ ID NO:130.

Human 8843 contains the following regions or other structural features: a dual specificity phosphatase domain (PFAM Accession Number PF00782) located at about amino acid residues 37 to 185 of SEQ ID NO:130; a tyrosine specific protein phosphatase active site signature (Prosite PS00383), also termed “C—X ₅—R” motif (SEQ ID NO:135), located at about amino acid residues 130 to 142 of SEQ ID NO:130, including an active site cysteine at about amino acid 132 of SEQ ID NO:130, and an active site arginine at about amino acid 138 of SEQ ID NO:130; a dual specificity phosphatase extended active site signature (VXVHCXXGXSRSXTXXXAY[LI]M; SEQ ID NO:136; Muda et al. (1996) J Biol Chem 271:27205) locate at about amino acid residues 128 to 158 of SEQ ID NO:130; a VH1-like dual specificity phosphatase loop located at about amino acid residues 106 to 110 of SEQ ID NO:130, include a conserved general acid, aspartic acid at about residue 109 of SEQ ID NO:130; one predicted N-glycosylation site (PS00001) at about amino acids 82 to 85 of SEQ ID NO:130; one predicted protein kinase C phosphorylation sites (PS00005) at about amino acids 187 to 189 of SEQ ID NO:130; four predicted casein kinase II phosphorylation sites (PS00006) located at about amino acids 70 to 73, 83 to 86, 98 to 101, and 154 to 157 of SEQ ID NO:130; and one predicted N-myristylation sites (PS00008) from about amino acid 114 to 119 of SEQ ID NO:130.

A hydropathy plot of human 8843 was performed. Polypeptides of the invention include fragments which include: all or part of a hydrophobic sequence, e.g., the sequence from about amino acid 9 to 25, from about 45 to 52, and from about 142 to 150, of SEQ ID NO:130; all or part of a hydrophilic sequence, e.g., the sequence of from about amino acid 27 to 36, from about 70 to 78, and from about 151 to 160, of SEQ ID NO:130; a sequence which includes a Cys, or a glycosylation site.

The 8843 protein contains a significant number of structural characteristics in common with members of the dual specificity phosphatase family.

Dual specificity phosphatase proteins are characterized by a common fold. Dual specificity phosphatases are exemplified by the VH1 or vaccinia virus late H1 gene protein, which hydrolyzes both phosphotyrosine, phosphothreonine, and phosphoserine. VH1 catalytic activity is required for viral replication. A human homolog of VH1, VHR, has been identified. The three dimensional structure of this family is based on models from x-ray crystallographic data of protein tyrosine phosphatases, and human VHR. The VHR structure includes a core domain consisting of a five-stranded mixed β-sheet and six α-helices. This structure closely superimposes on the structure of phosphotyrosine protein phosphatases. However, dual specificity phosphatases lack the KNRY motif, and the N-terminal structures of tyrosine protein phosphatases which endow these enzymes with a deep active site specific for aryl phosphates. Thus, dual specificity phosphatases have a shallower active site relative to tyrosine protein phosphatases and can accommodate phosphoserine and phosphothreonine substrates. Even so dual specificity phosphatases can have a greater than 50-fold faster rate of phosphatase activity for phosphotyrosine substrates than phosphothreonine or phosphoserine substrates.

Similar to the broader class of phosphatases, dual specificity phosphatases have a highly conserved active site including three catalytic residues, a cysteine, an arginine, and an aspartic acid. The active site cysteine and arginine are found in the “C—X ₅—R” motif of the tyrosine phosphatase signature (Prosite PS00383; SEQ ID NO:135). This motif forms a binding pocket for three of the phosphate oxyanions. The cysteine acts as a nucleophile to accept the PO₃group. The reaction transiently generates a phospho-cysteine intermediate before the phosphate is transferred to water. The active site arginine stabilizes the transition-state by hydrogen bonding to phosphate oxygens. In addition the histidine preceding the active site cysteine and the serine or threonine following the active site arginine are responsible for lowering the pK_aof the cysteine to stabilize a negative charge on the cysteine. The active site aspartic acid accelerates the reaction by donating a protein to generate an uncharged hydroxyl (for a review, see Fauman and Saper (1996) Trends in Biochem. 21:412).

An 8843 polypeptide can include a “dual specificity phosphatase catalytic domain” or regions homologous with a “dual specificity phosphatase domain”.

As used herein, the term “dual specificity phosphatase domain” includes an amino acid sequence of about 80 to 220 amino acids, more preferably about 100 to 180 amino acid residues, or about 130 to 160 amino acid residues in length and having a bit score for the alignment of the sequence to the dual specificity phosphatase domain (HMM) of at least 10, preferably 15, and more preferably 20. The dual specificity phosphatase catalytic domain (HMM) has been assigned the PFAM Accession Number PF00782. The dual specificity phosphatase domain (amino acids 37 to 185 of SEQ ID NO:130) of human 8843 aligns with a consensus amino acid sequence (SEQ ID NO:132) derived from a hidden Markov model, and with a consensus amino acid sequence (SEQ ID NO:133) derived from the SMART domain HMM model.

A dual specificity phosphatase domain preferably includes a perfect match to the Prosite tyrosine specific protein phosphatase active site signature (PS00383; [LIVMF]-H-C-x(2)-G-x(3)-[STC]-[STAGP]-x-[LIVMFY], wherein X is any amino acid and a number in parenthesis indicates the amino acid pattern is repeated that number of times; SEQ ID NO:134). Even more preferably, a dual specificity phosphatase includes the extended active site signature (VXVHCXXGXSRSXTXXXAY[LI]M; SEQ ID NO:136; Muda et al. (1996) J Biol Chem 271:27205). A dual specificity phosphatase domain also includes the conserved active site residues cysteine, arginine, and aspartic acid. The aspartic acid is preferably located in a loop region N-terminal to the active site signature.

In a preferred embodiment 8843 polypeptide or protein has a “dual specificity phosphatase domain” or a region which includes at least about 80 to 220 more preferably about 100 to 180 or 130 to 160, or about 148 amino acid residues in length and has at least about 50%, 60%, 70%, 80%, 90%, 95%, 99%, or 100% homology with a “dual specificity phosphatase domain,” e.g., the dual specificity phosphatase domain of human 8843 (e.g., residues 37 to 185 of SEQ ID NO:130). In a preferred embodiment, the 8834 polypeptide has a tyrosine specific protein phosphatase active site signature located at about amino acids 130 to 142 of SEQ ID NO:130. The 8834 polypeptide also preferable has a conserved active site serine at about amino acid residue 132 of SEQ ID NO:130, a conserved active site arginine at about amino acid residue 138 of SEQ ID NO:130, and a conserved active site aspartic acid at about amino acid residue 109 of SEQ ID NO:130. Preferably, the active site aspartic acid is in an mobile loop, approximately 20 to 30, or preferably, 20 to 25 amino acids N-terminal to the active site cysteine.

To identify the presence of a “dual specificity phosphatase” domain in an 8843 protein sequence, and make the determination that a polypeptide or protein of interest has a particular profile, the amino acid sequence of the protein can be searched against a database of HMMs (e.g., the Pfam database, release 2.1) using the default parameters. For example, the hmmsf program, which is available as part of the HMMER package of search programs, is a family specific default program for MILPAT0063 and a score of 15 is the default threshold score for determining a hit. Alternatively, the threshold score for determining a hit can be lowered (e.g., to 8 bits). A description of the Pfam database can be found in Sonhammer et al. (1997) Proteins 28(3):405-420 and a detailed description of HMMs can be found, for example, in Gribskov et al.(1990) Meth. Enzymol. 183:146-159; Gribskov et al.(1987) Proc. Natl. Acad. Sci. USA 84:4355-4358; Krogh et al.(1994) J. Mol. Biol. 235:1501-1531; and Stultz et al.(1993) Protein Sci. 2:305-314, the contents of which are incorporated herein by reference. A search was performed against the HMM database resulting in the identification of a “dual specificity phosphatase” domain in the amino acid sequence of human 8843 at about residues 37 to 185 of SEQ ID NO:130.

An 8843 molecule can further include: preferably at least one N-glycosylation site; preferably at least one protein kinase C phosphorylation sites; at least one, two, three, and preferably four casein kinase II phosphorylation sites; and preferably at least one N-myristylation site.

As used herein, a “8843 activity”, “biological activity of 8843” or “functional activity of 8843”, refers to an activity exerted by an 8843 protein, polypeptide or nucleic acid molecule on e.g., an 8843-responsive cell or on an 8843 substrate, e.g., a protein substrate, as determined in vivo or in vitro. In one embodiment, an 8843 activity is a direct activity, such as an association with an 8843 target molecule. A “target molecule” or “binding partner” is a molecule with which an 8843 protein binds or interacts in nature an 8843 -activity can also be an indirect activity, e.g., a cellular signaling activity mediated by interaction of the 8843 protein with an 8843 receptor. Based on the above-described sequence similarities, the 8843 molecules of the present invention are predicted to have similar biological activities as dual specificity phosphatase family members. For example, the 8843 proteins of the present invention can have one or more of the following activities: (1) catalyzing the removal of a phosphate group attached to a tyrosine residue in a protein; (2) catalyzing the removal of a phosphate group attached to a serine or threonine residue in a protein; (3) modulating an intracellular signaling pathway, e.g., a MAP kinase or ERK kinase pathway; (4) modulating cell differentiation, e.g., differentiation of erythroid progenitor cells, such as, CD34+ progenitors; (5) modulating cell proliferation, e.g., proliferation erythroid progenitor cells; (6) inactivating cell surface growth factor receptors, e.g., tyrosine kinase receptors; or (7) modulating apoptosis, of a cell, e.g., a leukemic cell, (e.g., an erythroleukemia cell).

As TaqMan analysis shows, 8843 mRNA is found in hematopoietic cells, and in particular, in erythroid cell lineages. The molecules of the invention can be used to develop novel agents or compounds to treat and/or diagnose disorders involving aberrant activities of those cells e.g., hematopoietic and, in particular, erythroid disorders, as described below. For example, an 8843 polypeptide is expressed in CD34 positive cells, e.g., mobilized peripheral blood CD34+ cells, normal adult bone marrow CD34+ cells, cord blood CD34+ cells, normal adult bone marrow CD34+ cells, G-CSF-treated bone marrow CD34+ cells, and fetal liver CD34+ cells; and erythroid progenitor cells, e.g., bone marrow glycophorin A positive cells and erythropoietin treated erythroid burst forming units (BFUs). 8843 mRNA is also expressed in hepatic cells, kidney, lung, and dermal cells, and thus diagnostic and therapeutic methods of using the molecules of the invention to treat/diagnose hepatic, kidney, lung, and dermal disorders are also contemplated by the present invention.

As used herein, the term “pluripotent hematopoietic stem cell” includes a cell that can give rise to a spleen colony forming unit (day 12 CFU-S), which, in turn, can give rise to progenitors of the granulocytic, monocytic, erythroid, megakaryocytic, and lymphoid lineages.

As used herein, a “CD34-positive cell” refers to a cell that expresses detectable levels of the CD34 antigen, preferably human CD34 antigen. The sequence for human CD34 is provided in SwissProt Accession Number P28906. The CD34 antigen is typically present on immature hematopoietic precursor cells and hematopoietic colony-forming cells in the bone marrow, including unipotent (CFU-GM, BFU-E) and pluripotent progenitors (CFU-GEMM, CFU-Mix and CFUJ-blast). The CD34 is also expressed on stromal cell precursors. Terminal deoxynucleotidyl transferase (TdT)-positive B- and T-lymphoid precursors in normal bone also are CD34+. The CD34 antigen is typically present on early myeloid cells that express the CD33 antigen, but lack the CD14 and CD15 antigens and on early erythroid cells that express the CD71 antigen and dimly express the CD45 antigen. The CD34 antigen is also found on capillary endothelial cells and approximately 1% of human thymocytes. Normal peripheral blood lymphocytes, monocytes, granulocytes and platelets do not express the CD34 antigen. CD34 antigen density is highest on early haematopoietic progenitor cells and decreases as the cells mature. The antigen is undetectably on fully differentiated haematopoietic cells. Approximately 60% of acute B-lymphoid leukemia's and acute myeloid leukemia express the CD34 antigen. The antigen is, not expressed on chronic lymphoid leukemia (B or T lineage) or lymphomas.

As the 8843 polypeptides of the invention may modulate 8843-mediated activities, they may be useful for developing novel diagnostic and therapeutic agents for 8843-mediated or related disorders, e.g., erythroid-associated disorders.

As used herein, the term “erythropoietin” or “EPO” refers to a glycoprotein produced in the kidney, which is the principal hormone responsible for stimulating red blood cell production (erythrogenesis). EPO stimulates the division and differentiation of committed erythroid progenitors in the bone marrow. Normal plasma erythropoietin levels range from 0.01 to 0.03 Units/mL, and can increase up to 100 to 1,000-fold during hypoxia: or anemia. Graber and Krantz, Ann. Rev. Med. 29:51 (1978); Eschbach and Adamson, Kidney Intl. 28:1 (1985). Recombinant human erythropoietin (rHuEpo or epoetin alfa) is commercially available as EPOGEN.RTM. (epoetin alfa, recombinant human erythropoietin) (Amgen Inc., Thousand Oaks, Calif.) and as PROCRIT.RTM. (epoetin alfa, recombinant human erythropoietin) (Ortho Biotech Inc., Raritan, N.J.).

Treatment, prevention and diagnosis of cancer or neoplastic disorders related to the erythroid lineage are also included in the present invention.

Tissue Distribution of 8843 mRNA

Endogenous human 8843 gene expression was determined using the Perkin-Elmer/ABI 7700 Sequence Detection System which employs TaqMan technology.

8843 mRNA levels were analyzed in a variety of samples of isolated and/or treated blood cells. High relative expression levels of 8843 mRNA, e.g., greater than 40 units, were observed for megakaryocytes, mast cells, blast forming units (BFU), especially BFUs treated with erythropoietin. Moderate relative expression levels of 8843 mRNA, between 10 and 40 units, were observed for multiple erythroid samples, and a subset of neutrophil samples.

High relative expression levels of 8843 mRNA, e.g., greater than 40 units, were observed for mobilized CD34+ peripheral blood cells (mBM), normal bone marrow CD34+ cells, as well as for glycophorin A (low levels) bone marrow cells (>60 units), which are erythroid progenitors. Moderate relative expression levels of 8843 mRNA, between 10 and 40 units, were observed for CD34+ cord blood cells, CD34+ fetal liver cells, and mobilized CD34+ bone marrow cells.

8843 mRNA expression levels were also monitored in other hematopoietic lineages and tissues. K582 cells, an erythroid/megakaryocyte cell line, and Hep3b cells had high 8843 expression levels relative to controls.

8843 mRNA expression was also determined for mRNA derived from lung, fetal liver, and other tissues.

Definitions

The 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein, fragments thereof, and derivatives and other variants of the sequence in SEQ ID NO:2, 5, 8, 11, 14, 17, 54, 62, 68, 79, 89, 101, 114, 123 or 130 thereof are collectively referred to as “polypeptides or proteins of the invention” or “26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 polypeptides or proteins”. Nucleic acid molecules encoding such polypeptides or proteins are collectively referred to as “nucleic acids of the invention” or “26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 nucleic acids.”

As used herein, the term “nucleic acid molecule” includes DNA molecules (e.g., a cDNA or genomic DNA) and RNA molecules (e.g., an mRNA) and analogs of the DNA or RNA generated, e.g., by the use of nucleotide analogs. The nucleic acid molecule can be single-stranded or double-stranded, but preferably is double-stranded DNA.

The term “isolated or purified nucleic acid molecule” includes nucleic acid molecules which are separated from other nucleic acid molecules which are present in the natural source of the nucleic acid. For example, with regards to genomic DNA, the term “isolated” includes nucleic acid molecules which are separated from the chromosome with which the genomic DNA is naturally associated. Preferably, an “isolated” nucleic acid is free of sequences which naturally flank the nucleic acid (i.e., sequences located at the 5′ and/or 3′ ends of the nucleic acid) in the genomic DNA of the organism from which the nucleic acid is derived. For example, in various embodiments, the isolated nucleic acid molecule can contain less than about 5 kb, 4 kb, 3 kb, 2 kb, 1 kb, 0.5 kb or 0.1 kb of 5′ and/or 3′ nucleotide sequences which naturally flank the nucleic acid molecule in genomic DNA of the cell from which the nucleic acid is derived. Moreover, an “isolated” nucleic acid molecule, such as a cDNA molecule, can be substantially free of other cellular material or culture medium when produced by recombinant techniques, or substantially free of chemical precursors or other chemicals when chemically synthesized.

As used herein, the term “hybridizes under low stringency, medium stringency, high stringency, or very high stringency conditions” describes conditions for hybridization and washing. Guidance for performing hybridization reactions can be found in Current Protocols in Molecular Biology (1989) John Wiley & Sons, N.Y., 6.3.1-6.3.6, which is incorporated by reference. Aqueous and nonaqueous methods are described in that reference and either can be used. Specific hybridization conditions referred to herein are as follows: 1) low stringency hybridization conditions in 6× sodium chloride/sodium citrate (SSC) at about 45° C., followed by two washes in 0.2×SSC, 0.1% SDS at least at 50° C. (the temperature of the washes can be increased to 55° C. for low stringency conditions); 2) medium stringency hybridization conditions in 6×SSC at about 45° C., followed by one or more washes in 0.2×SSC, 0.1% SDS at 60° C.; 3). high stringency hybridization conditions in 6×SSC at about 45° C., followed by one or more washes in 0.2×SSC, 0.1% SDS at 65° C.; and preferably 4) very high stringency hybridization conditions are 0.5M sodium phosphate, 7% SDS at 65° C., followed by one or more washes at 0.2×SSC, 1% SDS at 65° C. Very high stringency conditions (4) are the preferred conditions and the ones that should be used unless otherwise specified.

As used herein, a “naturally-occurring” nucleic acid molecule refers to an RNA or DNA molecule having a nucleotide sequence that occurs in nature (e.g., encodes a natural protein).

As used herein, the terms “gene” and “recombinant gene” refer to nucleic acid molecules which include an open reading frame encoding a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein, preferably a mammalian 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein, and can further include non-coding regulatory sequences, and introns.

An “isolated” or “purified” polypeptide or protein is substantially free of cellular material or other contaminating proteins from the cell or tissue source from which the protein is derived, or substantially free from chemical precursors or other chemicals when chemically synthesized. In one embodiment, the language “substantially free” means preparation of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein having less than about 30%, 20%, 10% and more preferably 5% (by dry weight), of non-26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein (also referred to herein as a “contaminating protein”), or of chemical precursors or non-26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 chemicals. When the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein or biologically active portion thereof is recombinantly produced, it is also preferably substantially free of culture medium, i.e., culture medium represents less than about 20%, more preferably less than about 10%, and most preferably less than about 5% of the volume of the protein preparation. The invention includes isolated or purified preparations of at least 0.01, 0.1, 1.0, and 10 milligrams in dry weight.

A “non-essential” amino acid residue is a residue that can be altered from the wild-type sequence of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 (e.g., the sequence of SEQ ID NO:1, 3, 4, 6, 7, 9, 10, 12, 13, 15, 16, 18, 53, 55, 61, 63, 67, 69, 78, 80, 88, 90, 100, 102, 113, 115, 122, 124, 129 or 131) without abolishing or more preferably, without substantially altering a biological activity, whereas an “essential” amino acid residue results in such a change. For example, amino acid residues that are conserved among the polypeptides of the present invention, e.g., those present in the conserved domains, are predicted to be particularly unamenable to alteration.

A “conservative amino acid substitution” is one in which the amino acid residue is replaced with an amino acid residue having a similar side chain. Families of amino acid residues having similar side chains have been defined in the art. These families include amino acids with basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan), beta-branched side chains (e.g., threonine, valine, isoleucine) and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine). Thus, a predicted nonessential amino acid residue in a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein is preferably replaced with another amino acid residue from the same side chain family. Alternatively, in another embodiment, mutations can be introduced randomly along all or part of a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 coding sequence, such as by saturation mutagenesis, and the resultant mutants can be screened for 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 biological activity to identify mutants that retain activity. Following mutagenesis of SEQ ID NO:1, 3, 4, 6, 7, 9, 10, 12, 13, 15, 16, 18, 53, 55, 61, 63, 67, 69, 78, 80, 88, 90, 100, 102, 113, 115, 122, 124, 129 or 131, the encoded protein can be expressed recombinantly and the activity of the protein can be determined.

As used herein, a “biologically active portion” of a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein includes a fragment of a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein which participates in an interaction between a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 molecule and a non-26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 molecule. Biologically active portions of a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein include peptides comprising amino acid sequences sufficiently homologous to or derived from the amino acid sequence of the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein, e.g., the amino acid sequence shown in SEQ ID NO:2, 5, 8, 11, 14, 17, 54, 62, 68, 79, 89, 101, 114, 123 or 130, which include fewer amino acids than the full length 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein, and exhibit at least one activity of a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein. Typically, biologically active portions comprise a domain or motif with at least one activity of the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein. A biologically active portion of a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein can be a polypeptide which is, for example, 10, 25, 50, 100, 200 or more amino acids in length. Biologically active portions of a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein can be used as targets for developing agents which modulate a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 mediated activity.

Calculations of homology or sequence identity (the terms “homology” and “identity” are used interchangeably herein) between sequences are performed as follows:

To determine the percent identity of two amino acid sequences, or of two nucleic acid sequences, the sequences are aligned for optimal comparison purposes (e.g., gaps can be introduced in one or both of a first and a second amino acid or nucleic acid sequence for optimal alignment and non-homologous sequences can be disregarded for comparison purposes). In a preferred embodiment, the length of a reference sequence aligned for comparison purposes is at least 30%, preferably at least 40%, more preferably at least 50%, even more preferably at least 60%, and even more preferably at least 70%, 80%, 90%, 100% of the length of the reference sequence. The amino acid residues or nucleotides at corresponding amino acid positions or nucleotide positions are then compared. When a position in the first sequence is occupied by the same amino acid residue or nucleotide as the corresponding position in the second sequence, then the molecules are identical at that position (as used herein amino acid or nucleic acid “identity” is equivalent to amino acid or nucleic acid “homology”). The percent identity between the two sequences is a function of the number of identical positions shared by the sequences, taking into account the number of gaps, and the length of each gap, which need to be introduced for optimal alignment of the two sequences.

The comparison of sequences and determination of percent identity between two sequences can be accomplished using a mathematical algorithm. In a preferred embodiment, the percent identity between two amino acid sequences is determined using the Needleman and Wunsch (1970) J. Mol. Biol. 48:444-453 algorithm which has been incorporated into the GAP program in the GCG software package using either a Blossum 62 matrix or a PAM250 matrix, and a gap weight of 16, 14, 12, 10, 8, 6, or 4 and a length weight of 1, 2, 3, 4, 5, or 6. In yet another preferred embodiment, the percent identity between two nucleotide sequences is determined using the GAP program in the GCG software package using a NWSgapdna.CMP matrix and a gap weight of 40, 50, 60, 70, or 80 and a length weight of 1, 2, 3, 4, 5, or 6. A particularly preferred set of parameters (and the one that should be used if the practitioner is uncertain about what parameters should be applied to determine if a molecule is within a sequence identity or homology limitation of the invention) are a Blossum 62 scoring matrix with a gap penalty of 12, a gap extend penalty of 4, and a frameshift gap penalty of 5.

The percent identity between two amino acid or nucleotide sequences can be determined using the algorithm of Meyers and Miller ((1989) CABIOS, 4:11-17) which has been incorporated into the ALIGN program (version 2.0), using a PAM120 weight residue table, a gap length penalty of 12 and a gap penalty of 4.

The nucleic acid and protein sequences described herein can be used as a “query sequence” to perform a search against public databases to, for example, identify other family members or related sequences. Such searches can be performed using the NBLAST and XBLAST programs (version 2.0) of Altschul et al. (1990) J. Mol. Biol. 215:403-10. BLAST nucleotide searches can be performed with the NBLAST program, score=100, wordlength=12 to obtain nucleotide sequences homologous to 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 nucleic acid molecules of the invention. BLAST protein searches can be performed with the XBLAST program, score=50, wordlength=3 to obtain amino acid sequences homologous to 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein molecules of the invention. To obtain gapped alignments for comparison purposes, Gapped BLAST can be utilized as described in Altschul et al., (1997) Nucleic Acids Res. 25:3389-3402. When utilizing BLAST and Gapped BLAST programs, the default parameters of the respective programs (e.g., XBLAST and NBLAST) can be used.

Particular 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 polypeptides of the present invention have an amino acid sequence substantially identical to the amino acid sequence of SEQ ID NO:2, 5, 8, 11, 14, 17, 54, 62, 68, 79, 89, 101, 114, 123 or 130. In the context of an amino acid sequence, the term “substantially identical” is used herein to refer to a first amino acid that contains a sufficient or minimum number of amino acid residues that are i) identical to, or ii) conservative substitutions of aligned amino acid residues in a second amino acid sequence such that the first and second amino acid sequences can have a common structural domain and/or common functional activity. For example, amino acid sequences that contain a common structural domain having at least about 60%, or 65% identity, likely 75% identity, more likely 85%, 90%. 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2, 5, 8, 11, 14, 17, 54, 62, 68, 79, 89, 101, 114, 123 or 130 are termed substantially identical.

In the context of nucleotide sequence, the term “substantially identical” is used herein to refer to a first nucleic acid sequence that contains a sufficient or minimum number of nucleotides that are identical to aligned nucleotides in a second nucleic acid sequence such that the first and second nucleotide sequences encode a polypeptide having common functional activity, or encode a common structural polypeptide domain or a common functional polypeptide activity. For example, nucleotide sequences having at least about 60%, or 65% identity, likely 75% identity, more likely 85%, 90%. 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:1, 3, 4, 6, 7, 9, 10, 12, 13, 15, 16, 18, 53, 55, 61, 63, 67, 69, 78, 80, 88, 90, 100, 102, 113, 115, 122, 124, 129 or 131 are termed substantially identical.

“Misexpression or aberrant expression”, as used herein, refers to a non-wild type pattern of gene expression, at the RNA or protein level. It includes: expression at non-wild type levels, i.e., over or under expression; a pattern of expression that differs from wild type in terms of the time or stage at which the gene is expressed, e.g., increased or decreased expression (as compared with wild type) at a predetermined developmental period or stage; a pattern of expression that differs from wild type in terms of decreased expression (as compared with wild type) in a predetermined cell type or tissue type; a pattern of expression that differs from wild type in terms of the splicing size, amino acid sequence, post-transitional modification, or biological activity of the expressed polypeptide; a pattern of expression that differs from wild type in terms of the effect of an environmental stimulus or extracellular stimulus on expression of the gene, e.g., a pattern of increased or decreased expression (as compared with wild type) in the presence of an increase or decrease in the strength of the stimulus.

“Subject”, as used herein, can refer to a mammal, e.g., a human, or to an experimental or animal or disease model. The subject can also be a non-human animal, e.g., a horse, cow, goat, or other domestic animal.

A “purified preparation of cells”, as used herein, refers to, in the case of plant or animal cells, an in vitro preparation of cells and not an entire intact plant or animal. In the case of cultured cells or microbial cells, it consists of a preparation of at least 10% and more preferably 50% of the subject cells.

As used herein, cellular proliferative and/or differentiative disorders include cancer, e.g., carcinoma, sarcoma, metastatic disorders or hematopoietic neoplastic disorders, e.g., leukemias. A metastatic tumor can arise from a multitude of primary tumor types, including but not limited to those of prostate, colon, lung, breast and liver origin.

As used herein, the term “cancer” (also used interchangeably with the terms, “hyperproliferative” and “neoplastic”) refers to cells having the capacity for autonomous growth, i.e., an abnormal state or condition characterized by rapidly proliferating cell growth. Cancerous disease states may be categorized as pathologic, i.e., characterizing or constituting a disease state, e.g., malignant tumor growth, or may be categorized as non-pathologic, i.e., a deviation from normal but not associated with a disease state, e.g., cell proliferation associated with wound repair. The term is meant to include all types of cancerous growths or oncogenic processes, metastatic tissues or malignantly transformed cells, tissues, or organs, irrespective of histopathologic type or stage of invasiveness. The term “cancer” includes malignancies of the various organ systems, such as those affecting lung, breast, thyroid, lymphoid, gastrointestinal, and genito-urinary tract, as well as adenocarcinomas which include malignancies such as most colon cancers, renal-cell carcinoma, prostate cancer and/or testicular tumors, non-small cell carcinoma of the lung, cancer of the small intestine and cancer of the esophagus. The term “carcinoma” is art recognized and refers to malignancies of epithelial or endocrine tissues including respiratory system carcinomas, gastrointestinal system carcinomas, genitourinary system carcinomas, testicular carcinomas, breast carcinomas, prostatic carcinomas, endocrine system carcinomas, and melanomas. Exemplary carcinomas include those forming from tissue of the cervix, lung, prostate, breast, head and neck, colon and ovary. The term “carcinoma” also includes carcinosarcomas, e.g., which include malignant tumors composed of carcinomatous and sarcomatous tissues. An “adenocarcinoma” refers to a carcinoma derived from glandular tissue or in which the tumor cells form recognizable glandular structures. The term “sarcoma” is art recognized and refers to malignant tumors of mesenchymal derivation.

Examples of cellular proliferative and/or differentiative disorders of the lung include, but are not limited to, tumors such as bronchogenic carcinoma, including paraneoplastic syndromes, bronchioloalveolar carcinoma, neuroendocrine tumors, such as bronchial carcinoid, miscellaneous tumors, metastatic tumors, and pleural tumors, including solitary fibrous tumors (pleural fibroma) and malignant mesothelioma.

Examples of cellular proliferative and/or differentiative disorders of the breast include, but are not limited to, proliferative breast disease including, e.g., epithelial hyperplasia, sclerosing adenosis, and small duct papillomas; tumors, e.g., stromal tumors such as fibroadenoma, phyllodes tumor, and sarcomas, and epithelial tumors such as large duct papilloma; carcinoma of the breast including in situ (noninvasive) carcinoma that includes ductal carcinoma in situ (including Paget's disease) and lobular carcinoma in situ, and invasive (infiltrating) carcinoma including, but not limited to, invasive ductal carcinoma, invasive lobular carcinoma, medullary carcinoma, colloid (mucinous) carcinoma, tubular carcinoma, and invasive papillary carcinoma, and miscellaneous malignant neoplasms. Disorders in the male breast include, but are not limited to, gynecomastia and carcinoma.

Examples of cellular proliferative and/or differentiative disorders involving the colon include, but are not limited to, tumors of the colon, such as non-neoplastic polyps, adenomas, familial syndromes, colorectal carcinogenesis; colorectal carcinoma, and carcinoid tumors.

Examples of cancers or neoplastic conditions, in addition to the ones described above, include, but are not limited to, a fibrosarcoma, myosarcoma, liposarcoma, chondrosarcoma, osteogenic sarcoma, chordoma, angiosarcoma, endotheliosarcoma, lymphangiosarcoma, lymphangioendotheliosarcoma, synovioma, mesothelioma, Ewing's tumor, leiomyosarcoma, rhabdomyosarcoma, gastric cancer, esophageal cancer, rectal cancer, pancreatic cancer, ovarian cancer, prostate cancer, uterine cancer, cancer of the head and neck, skin cancer, brain cancer, squamous cell carcinoma, sebaceous gland carcinoma, papillary carcinoma, papillary adenocarcinoma, cystadenocarcinoma, medullary carcinoma, bronchogenic carcinoma, renal cell carcinoma, hepatoma, bile duct carcinoma, choriocarcinoma, seminoma, embryonal carcinoma, Wilm's tumor, cervical cancer, testicular cancer, small cell lung carcinoma, non-small cell lung carcinoma, bladder carcinoma, epithelial carcinoma, glioma, astrocytoma, medulloblastoma, craniopharyngioma, ependymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, meningioma, melanoma, neuroblastoma, retinoblastoma, leukemia, lymphoma, or Kaposi sarcoma.

Proliferative disorders include hematopoietic neoplastic disorders. As used herein, the term “hematopoietic neoplastic disorders” includes diseases involving hyperplastic/neoplastic cells of hematopoietic origin, e.g., arising from myeloid, lymphoid or erythroid lineages, or precursor cells thereof. Preferably, the diseases arise from poorly differentiated acute leukemias, e.g., erythroblastic leukemia and acute megakaryoblastic leukemia. Additional exemplary myeloid disorders include, but are not limited to, acute promyeloid leukemia (APML), acute myelogenous leukemia (AML) and chronic myelogenous leukemia (CML) (reviewed in Vaickus (1991) Crit Rev. in Oncol./Hemotol. 11:267-97); lymphoid malignancies include, but are not limited to acute lymphoblastic leukemia (ALL) which includes B-lineage ALL and T-lineage ALL, chronic lymphocytic leukemia (CLL), prolymphocytic leukemia (PLL), hairy cell leukemia (HLL) and Waldenstrom's macroglobulinemia (WM). Additional forms of malignant lymphomas include, but are not limited to non-Hodgkin lymphoma and variants thereof, peripheral T cell lymphomas, adult T cell leukemia/lymphoma (ATL), cutaneous T-cell lymphoma (CTCL), large granular lymphocytic leukemia (LGF), Hodgkin's disease and Reed-Sternberg disease.

As used herein, disorders of the breast include, but are not limited to, disorders of development; inflammations, including but not limited to, acute mastitis, periductal mastitis, periductal mastitis (recurrent subareolar abscess, squamous metaplasia of lactiferous ducts), mammary duct ectasia, fat necrosis, granulomatous mastitis, and pathologies associated with silicone breast implants; fibrocystic changes; proliferative breast disease including, but not limited to, epithelial hyperplasia, sclerosing adenosis, and small duct papillomas; tumors including, but not limited to, stromal tumors such as fibroadenoma, phyllodes tumor, and sarcomas, and epithelial tumors such as large duct papilloma; carcinoma of the breast including in situ (noninvasive) carcinoma that includes ductal carcinoma in situ (including Paget's disease) and lobular carcinoma in situ, and invasive (infiltrating) carcinoma including, but not limited to, invasive ductal carcinoma, no special type, invasive lobular carcinoma, medullary carcinoma, colloid (mucinous) carcinoma, tubular carcinoma, and invasive papillary carcinoma, and miscellaneous malignant neoplasms. Disorders in the male breast include, but are not limited to, gynecomastia and carcinoma.

As used herein, disorders involving the colon include, but are not limited to, congenital anomalies, such as atresia and stenosis, Meckel diverticulum, congenital aganglionic megacolon-Hirschsprung disease; enterocolitis, such as diarrhea and dysentery, infectious enterocolitis, including viral gastroenteritis, bacterial enterocolitis, necrotizing enterocolitis, antibiotic-associated colitis (pseudomembranous colitis), and collagenous and lymphocytic colitis, miscellaneous intestinal inflammatory disorders, including parasites and protozoa, acquired immunodeficiency syndrome, transplantation, drug-induced intestinal injury, radiation enterocolitis, neutropenic colitis (typhlitis), and diversion colitis; idiopathic inflammatory bowel disease, such as Crohn disease and ulcerative colitis; tumors of the colon, such as non-neoplastic polyps, adenomas, familial syndromes, colorectal carcinogenesis, colorectal carcinoma, and carcinoid tumors.

As used herein, disorders involving the kidney (or renal disorders) include, but are not limited to, congenital anomalies including, but not limited to, cystic diseases of the kidney, that include but are not limited to, cystic renal dysplasia, autosomal dominant (adult) polycystic kidney disease, autosomal recessive (childhood) polycystic kidney disease, and cystic diseases of renal medulla, which include, but are not limited to, medullary sponge kidney, and nephronophthisis-uremic medullary cystic disease complex, acquired (dialysis-associated) cystic disease, such as simple cysts; glomerular diseases including pathologies of glomerular injury that include, but are not limited to, in situ immune complex deposition, that includes, but is not limited to, anti-GBM nephritis, Heymann nephritis, and antibodies against planted antigens, circulating immune complex nephritis, antibodies to glomerular cells, cell-mediated immunity in glomerulonephritis, activation of alternative complement pathway, epithelial cell injury, and pathologies involving mediators of glomerular injury including cellular and soluble mediators, acute glomerulonephritis, such as acute proliferative (poststreptococcal, postinfectious) glomerulonephritis, including but not limited to, poststreptococcal glomerulonephritis and nonstreptococcal acute glomerulonephritis, rapidly progressive (crescentic) glomerulonephritis, nephrotic syndrome, membranous glomerulonephritis (membranous nephropathy), minimal change disease (lipoid nephrosis), focal segmental glomerulosclerosis, membranoproliferative glomerulonephritis, IgA nephropathy (Berger disease), focal proliferative and necrotizing glomerulonephritis (focal glomerulonephritis), hereditary nephritis, including but not limited to, Alport syndrome and thin membrane disease (benign familial hematuria), chronic glomerulonephritis, glomerular lesions associated with systemic disease, including but not limited to, systemic lupus erythematosus, Henoch-Schonlein purpura, bacterial endocarditis, diabetic glomerulosclerosis, amyloidosis, fibrillary and immunotactoid glomerulonephritis, and other systemic disorders; diseases affecting tubules and interstitium, including acute tubular necrosis and tubulointerstitial nephritis, including but not limited to, pyelonephritis and urinary tract infection, acute pyelonephritis, chronic pyelonephritis and reflux nephropathy, and tubulointerstitial nephritis induced by drugs and toxins, including but not limited to, acute drug-induced interstitial nephritis, analgesic abuse nephropathy, nephropathy associated with nonsteroidal anti-inflammatory drugs, and other tubulointerstitial diseases including, but not limited to, urate nephropathy, hypercalcemia and nephrocalcinosis, and multiple myeloma; diseases of blood vessels including benign nephrosclerosis, malignant hypertension and accelerated nephrosclerosis, renal artery stenosis, and thrombotic microangiopathies including, but not limited to, classic (childhood) hemolytic-uremic syndrome, adult hemolytic-uremic syndrome/thrombotic thrombocytopenic purpura, idiopathic HUS/TTP, and other vascular disorders including, but not limited to, atherosclerotic ischemic renal disease, atheroembolic renal disease, sickle cell disease nephropathy, diffuse cortical necrosis, and renal infarcts; urinary tract obstruction (obstructive uropathy); urolithiasis (renal calculi, stones); and tumors of the kidney including, but not limited to, benign tumors, such as renal papillary adenoma, renal fibroma or hamartoma (renomedullary interstitial cell tumor), angiomyolipoma, and oncocytoma, and malignant tumors, including renal cell carcinoma (hypernephroma, adenocarcinoma of kidney), which includes urothelial carcinomas of renal pelvis.

Examples of disorders of the lung include, but are not limited to, congenital anomalies; atelectasis; diseases of vascular origin, such as pulmonary congestion and edema, including hemodynamic pulmonary edema and edema caused by microvascular injury, adult respiratory distress syndrome (diffuse alveolar damage), pulmonary embolism, hemorrhage, and infarction, and pulmonary hypertension and vascular sclerosis; chronic obstructive pulmonary disease, such as emphysema, chronic bronchitis, bronchial asthma, and bronchiectasis; diffuse interstitial (infiltrative, restrictive) diseases, such as pneumoconioses, sarcoidosis, idiopathic pulmonary fibrosis, desquamative interstitial pneumonitis, hypersensitivity pneumonitis, pulmonary eosinophilia (pulmonary infiltration with eosinophilia), Bronchiolitis obliterans-organizing pneumonia, diffuse pulmonary hemorrhage syndromes, including Goodpasture syndrome, idiopathic pulmonary hemosiderosis and other hemorrhagic syndromes, pulmonary involvement in collagen vascular disorders, and pulmonary alveolar proteinosis; complications of therapies, such as drug-induced lung disease, radiation-induced lung disease, and lung transplantation; tumors, such as bronchogenic carcinoma, including paraneoplastic syndromes, bronchioloalveolar carcinoma, neuroendocrine tumors, such as bronchial carcinoid, miscellaneous tumors, and metastatic tumors; pathologies of the pleura, including inflammatory pleural effusions, noninflammatory pleural effusions, pneumothorax, and pleural tumors, including solitary fibrous tumors (pleural fibroma) and malignant mesothelioma.

As used herein disorders involving the pancreas include those of the exocrine pancreas such as congenital anomalies, including but not limited to, ectopic pancreas; pancreatitis, including but not limited to, acute pancreatitis; cysts, including but not limited to, pseudocysts; tumors, including but not limited to, cystic tumors and carcinoma of the pancreas; and disorders of the endocrine pancreas such as, diabetes mellitus; islet cell tumors, including but not limited to, insulinomas, gastrinomas, and other rare islet cell tumors.

As used herein, disorders involving the ovary include, for example, polycystic ovarian disease, Stein-leventhal syndrome, Pseudomyxoma peritonei and stromal hyperthecosis; ovarian tumors such as, tumors of coelomic epithelium, serous tumors, mucinous tumors, endometeriod tumors, clear cell adenocarcinoma, cystadenofibroma, brenner tumor, surface epithelial tumors; germ cell tumors such as mature (benign) teratomas, monoderrnal teratomas, immature malignant teratomas, dysgerminoma, endodermal sinus tumor, choriocarcinoma; sex cord-stomal tumors such as, granulosa-theca cell tumors, thecoma-fibromas, androblastomas, hill cell tumors, and gonadoblastoma; and metastatic tumors such as Krukenberg tumors.

Aberrant expression and/or activity of the molecules of the invention can mediate disorders associated with bone metabolism. “Bone metabolism” refers to direct or indirect effects in the formation or degeneration of bone structures, e.g., bone formation, bone resorption, etc., which can ultimately affect the concentrations in serum of calcium and phosphate. This term also includes activities mediated by the molecules of the invention in bone cells, e.g. osteoclasts and osteoblasts, that can in turn result in bone formation and degeneration. For example, molecules of the invention can support different activities of bone resorbing osteoclasts such as the stimulation of differentiation of monocytes and mononuclear phagocytes into osteoclasts. Accordingly, molecules of the invention that modulate the production of bone cells can influence bone formation and degeneration, and thus can be used to treat bone disorders. Examples of such disorders include, but are not limited to, osteoporosis, osteodystrophy, osteomalacia, rickets, osteitis fibrosa cystica, renal osteodystrophy, osteosclerosis, anti-convulsant treatment, osteopenia, fibrogenesis-imperfecta ossium, secondary hyperparathyrodism, hypoparathyroidism, hyperparathyroidism, cirrhosis, obstructive jaundice, drug induced metabolism, medullary carcinoma, chronic renal disease, rickets, sarcoidosis, glucocorticoid antagonism, malabsorption syndrome, steatorrhea, tropical sprue, idiopathic hypercalcemia and milk fever.

As used herein, “a prostate disorder” refers to an abnormal condition occurring in the male pelvic region characterized by, e.g., male sexual dysfunction and/or urinary symptoms. This disorder may be manifested in the form of genitourinary inflammation (e.g., inflammation of smooth muscle cells) as in several common diseases of the prostate including prostatitis, benign prostatic hyperplasia and cancer, e.g., adenocarcinoma or carcinoma, of the prostate.

Examples of immune, e.g., inflammatory, (e.g. respiratory inflammatory) disorders or diseases include, but are not limited to, autoimmune diseases (including, for example, diabetes mellitus, arthritis (including rheumatoid arthritis, juvenile rheumatoid arthritis, osteoarthritis, psoriatic arthritis), multiple sclerosis, encephalomyelitis, myasthenia gravis, systemic lupus erythematosis, autoimmune thyroiditis, dermatitis (including atopic dermatitis and eczematous dermatitis), psoriasis, Sjögren's Syndrome, inflammatory bowel disease, e.g. Crohn's disease and ulcerative colitis, aphthous ulcer, iritis, conjunctivitis, keratoconjunctivitis, asthma, allergic asthma, chronic obstructive pulmonary disease, cutaneous lupus erythematosus, scleroderma, vaginitis, proctitis, drug eruptions, leprosy reversal reactions, erythema nodosum leprosum, autoimmune uveitis, allergic encephalomyelitis, acute necrotizing hemorrhagic encephalopathy, idiopathic bilateral progressive sensorineural hearing loss, aplastic anemia, pure red cell anemia, idiopathic thrombocytopenia, polychondritis, Wegener's granulomatosis, chronic active hepatitis, Stevens-Johnson syndrome, idiopathic sprue, lichen planus, Graves' disease, sarcoidosis, primary biliary cirrhosis, uveitis posterior, and interstitial lung fibrosis), graft-versus-host disease, cases of transplantation, and allergy such as, atopic allergy.

As used herein, disorders involving the heart, or “cardiovascular disease” or a “cardiovascular disorder” includes a disease or disorder which affects the cardiovascular system, e.g., the heart, the blood vessels, and/or the blood. A cardiovascular disorder can be caused by an imbalance in arterial pressure, a malfunction of the heart, or an occlusion of a blood vessel, e.g., by a thrombus. A cardiovascular disorder includes, but is not limited to disorders such as arteriosclerosis, atherosclerosis, cardiac hypertrophy, ischemia reperfusion injury, restenosis, arterial inflammation, vascular wall remodeling, ventricular remodeling, rapid ventricular pacing, coronary microembolism, tachycardia, bradycardia, pressure overload, aortic bending, coronary artery ligation, vascular heart disease, valvular disease, including but not limited to, valvular degeneration caused by calcification, rheumatic heart disease, endocarditis, or complications of artificial valves; atrial fibrillation, long-QT syndrome, congestive heart failure, sinus node dysfunction, angina, heart failure, hypertension, atrial fibrillation, atrial flutter, pericardial disease, including but not limited to, pericardial effusion and pericarditis; cardiomyopathies, e.g., dilated cardiomyopathy or idiopathic cardiomyopathy, myocardial infarction, coronary artery disease, coronary artery spasm, ischemic disease, arrhythmia, sudden cardiac death, and cardiovascular developmental disorders (e.g., arteriovenous malformations, arteriovenous fistulae, raynaud's syndrome, neurogenic thoracic outlet syndrome, causalgia/reflex sympathetic dystrophy, hemangioma, aneurysm, cavernous angioma, aortic valve stenosis, atrial septal defects, atrioventricular canal, coarctation of the aorta, ebsteins anomaly, hypoplastic left heart syndrome, interruption of the aortic arch, mitral valve prolapse, ductus arteriosus, patent foramen ovale, partial anomalous pulmonary venous return, pulmonary atresia with ventricular septal defect, pulmonary atresia without ventricular septal defect, persistance of the fetal circulation, pulmonary valve stenosis, single ventricle, total anomalous pulmonary venous return, transposition of the great vessels, tricuspid atresia, truncus arteriosus, ventricular septal defects). A cardiovascular disease or disorder also can include an, endothelial cell disorder.

As used herein, disorders involving the brain include, but are not limited to, disorders involving neurons, and disorders involving glia, such as astrocytes, oligodendrocytes, ependymal cells, and microglia; cerebral edema, raised intracranial pressure and herniation, and hydrocephalus; malformations and developmental diseases, such as neural tube defects, forebrain anomalies, posterior fossa anomalies, and syringomyelia and hydromyelia; perinatal brain injury; cerebrovascular diseases, such as those related to hypoxia, ischemia, and infarction, including hypotension, hypoperfusion, and low-flow states—global cerebral ischemia and focal cerebral ischemia—infarction from obstruction of local blood supply, intracranial hemorrhage, including intracerebral (intraparenchymal) hemorrhage, subarachnoid hemorrhage and ruptured berry aneurysms, and vascular malformations, hypertensive cerebrovascular disease, including lacunar infarcts, slit hemorrhages, and hypertensive encephalopathy; infections, such as acute meningitis, including acute pyogenic (bacterial) meningitis and acute aseptic (viral) meningitis, acute focal suppurative infections, including brain abscess, subdural empyema, and extradural abscess, chronic bacterial meningoencephalitis, including tuberculosis and mycobacterioses, neurosyphilis, and neuroborreliosis (Lyme disease), viral meningoencephalitis, including arthropod-borne (Arbo) viral encephalitis, Herpes simplex virus Type 1, Herpes simplex virus Type 2, Varicella-zoster virus (Herpes zoster), cytomegalovirus, poliomyelitis, rabies, and human immunodeficiency virus 1, including HIV-1 meningoencephalitis (subacute encephalitis), vacuolar myelopathy, AIDS-associated myopathy, peripheral neuropathy, and AIDS in children, progressive multifocal leukoencephalopathy, subacute sclerosing panencephalitis, fungal meningoencephalitis, other infectious diseases of the nervous system; transmissible spongiform encephalopathies (prion diseases); demyelinating diseases, including multiple sclerosis, multiple sclerosis variants, acute disseminated encephalomyelitis and acute necrotizing hemorrhagic encephalomyelitis, and other diseases with demyelination; degenerative diseases, such as degenerative diseases affecting the cerebral cortex, including Alzheimer disease and Pick disease, degenerative diseases of basal ganglia and brain stem, including Parkinsonism, idiopathic Parkinson disease (paralysis agitans), progressive supranuclear palsy, corticobasal degenration, multiple system atrophy, including striatonigral degenration, Shy-Drager syndrome, and olivopontocerebellar atrophy, and Huntington disease; spinocerebellar degenerations, including spinocerebellar ataxias, including Friedreich ataxia, and ataxia-telanglectasia, degenerative diseases affecting motor neurons, including amyotrophic lateral sclerosis (motor neuron disease), bulbospinal atrophy (Kennedy syndrome), and spinal muscular atrophy; inborn errors of metabolism, such as leukodystrophies, including Krabbe disease, metachromatic leukodystrophy, adrenoleukodystrophy, Pelizaeus-Merzbacher disease, and Canavan disease, mitochondrial encephalomyopathies, including Leigh disease and other mitochondrial encephalomyopathies; toxic and acquired metabolic diseases, including vitamin deficiencies such as thiamine (vitamin B₁) deficiency and vitamin B₁₂deficiency, neurologic sequelae of metabolic disturbances, including hypoglycemia, hyperglycemia, and hepatic encephatopathy, toxic disorders, including carbon monoxide, methanol, ethanol, and radiation, including combined methotrexate and radiation-induced injury; tumors, such as gliomas, including astrocytoma, including fibrillary (diffuse) astrocytoma and glioblastoma multiforme, pilocytic astrocytoma, pleomorphic xanthoastrocytoma, and brain stem glioma, oligodendroglioma, and ependymoma and related paraventricular mass lesions, neuronal tumors, poorly differentiated neoplasms, including medulloblastoma, other parenchymal tumors, including primary brain lymphoma, germ cell tumors, and pineal parenchymal tumors, meningiomas, metastatic tumors, paraneoplastic syndromes, peripheral nerve sheath tumors, including schwannoma, neurofibroma, and malignant peripheral nerve sheath tumor (malignant schwannoma), and neurocutaneous syndromes (phakomatoses), including neurofibromotosis, including Type 1 neurofibromatosis (NF1) and TYPE 2 neurofibromatosis (NF2), tuberous sclerosis, and Von Hippel-Lindau disease.

As used herein, skeletal muscle disorders include, but are not limited to, muscular dystrophy (e.g., Duchenne muscular dystrophy, Becker muscular dystrophy, Emery-Dreifuss muscular dystrophy, limb-girdle muscular dystrophy, facioscapulohumeral muscular dystrophy, myotonic dystrophy, oculopharyngeal muscular dystrophy, distal muscular dystrophy, and congenital muscular dystrophy), motor neuron diseases (e.g., amyotrophic lateral sclerosis, infantile progressive spinal muscular atrophy, intermediate spinal muscular atrophy, spinal bulbar muscular atrophy, and adult spinal muscular atrophy), myopathies (e.g., inflammatory myopathies (e.g., dermatomyositis and polymyositis), myotonia congenita, paramyotonia congenita, central core disease, nemaline myopathy, myotubular myopathy, and periodic paralysis), tumors such as rhabdomyosarcoma, and metabolic diseases of muscle (e.g., phosphorylase deficiency, acid maltase deficiency, phosphofructokinase deficiency, debrancher enzyme deficiency, mitochondrial myopathy, carnitine deficiency, carnitine palmityl transferase deficiency, phosphoglycerate kinase deficiency, phosphoglycerate mutase deficiency, lactate dehydrogenase deficiency, and myoadenylate deaminase deficiency).

As used herein, an “endothelial cell disorder” includes a disorder characterized by aberrant, unregulated, or unwanted endothelial cell activity, e.g., proliferation, migration, angiogenesis, or vascularization; or aberrant expression of cell surface adhesion molecules or genes associated with angiogenesis, e.g., TIE-2, FLT and FLK. Endothelial cell disorders include tumorigenesis, tumor metastasis, psoriasis, diabetic retinopathy, endometriosis, Grave's disease, ischemic disease (e.g., atherosclerosis), and chronic inflammatory diseases (e.g., rheumatoid arthritis).

Disorders involving the liver (hepatic disorders) include, but are not limited to, hepatic injury; jaundice and cholestasis, such as bilirubin and bile formation; hepatic failure and cirrhosis, such as cirrhosis, portal hypertension, including ascites, portosystemic shunts, and splenomegaly; infectious disorders, such as viral hepatitis, including hepatitis A-E infection and infection by other hepatitis viruses, clinicopathologic syndromes, such as the carrier state, asymptomatic infection, acute viral hepatitis, chronic viral hepatitis, and fulminant hepatitis; autoimmune hepatitis; drug- and toxin-induced liver disease, such as alcoholic liver disease; inborn errors of metabolism and pediatric liver disease, such as hemochromatosis, Wilson disease, α ₁-antitrypsin deficiency, and neonatal hepatitis; primary bile acid malabsorption; intrahepatic biliary tract disease, such as secondary biliary cirrhosis, primary biliary cirrhosis, primary sclerosing cholangitis, and anomalies of the biliary tree; circulatory disorders, such as impaired blood flow into the liver, including hepatic artery compromise and portal vein obstruction and thrombosis, impaired blood flow through the liver, including passive congestion and centrilobular necrosis and peliosis hepatis, hepatic vein outflow obstruction, including hepatic vein thrombosis (Budd-Chiari syndrome) and veno-occlusive disease; hepatic disease associated with pregnancy, such as preeclampsia and eclampsia, acute fatty liver of pregnancy, and intrehepatic cholestasis of pregnancy; hepatic complications of organ or bone marrow transplantation, such as drug toxicity after bone marrow transplantation, graft-versus-host disease and liver rejection, and nonimmunologic damage to liver allografts; tumors and tumorous conditions, such as nodular hyperplasias, adenomas, and malignant tumors, including primary carcinoma of the liver and metastatic tumors.

Disorders which can be treated or diagnosed by methods described herein include, but are not limited to, disorders associated with an accumulation in the liver of fibrous tissue, such as that resulting from an imbalance between production and degradation of the extracellular matrix accompanied by the collapse and condensation of preexisting fibers. The methods described herein can be used to diagnose or treat hepatocellular necrosis or injury induced by a wide variety of agents including processes which disturb homeostasis, such as an inflammatory process, tissue damage resulting from toxic injury or altered hepatic blood flow, and infections (e.g., bacterial, viral and parasitic). For example, the methods can be used for the early detection of hepatic injury, such as portal hypertension or hepatic fibrosis. In addition, the methods can be employed to detect liver fibrosis attributed to inborn errors of metabolism, for example, fibrosis resulting from a storage disorder such as Gaucher's disease (lipid abnormalities) or a glycogen storage disease, A1-antitrypsin deficiency; a disorder mediating the accumulation (e.g., storage) of an exogenous substance, for example, hemochromatosis (iron-overload syndrome) and copper storage diseases (Wilson's disease), disorders resulting in the accumulation of a toxic metabolite (e.g., tyrosinemia, fructosemia and galactosemia) and peroxisomal disorders (e.g., Zellweger syndrome). Additionally, the methods described herein can be used for the early detection and treatment of liver injury associated with the administration of various chemicals or drugs, such as for example, methotrexate, isonizaid, oxyphenisatin, methyldopa, chlorpromazine, tolbutamide or alcohol, or which represents a hepatic manifestation of a vascular disorder such as obstruction of either the intrahepatic or extrahepatic bile flow or an alteration in hepatic circulation resulting, for example, from chronic heart failure, veno-occlusive disease, portal vein thrombosis or Budd-Chiari syndrome.

Additionally, the molecules of the invention can play an important role in the etiology of certain viral diseases, including but not limited to Hepatitis B, Hepatitis C and Herpes Simplex Virus (HSV). Modulators of the activity of the molecules of the invention could be used to control viral diseases. The modulators can be used in the treatment and/or diagnosis of viral infected tissue or virus-associated tissue fibrosis, especially liver and liver fibrosis. Also, such modulators can be used in the treatment and/or diagnosis of virus-associated carcinoma, especially hepatocellular cancer.

Disorders related to reduced platelet number, thrombocytopenia, include idiopathic thrombocytopenic purpura, including acute idiopathic thrombocytopenic purpura, drug-induced thrombocytopenia, HIV-associated thrombocytopenia, and thrombotic microangiopathies: thrombotic thrombocytopenic purpura and hemolytic-uremic syndrome.

As used herein, neurological disorders include disorders of the central nervous system (CNS) and the peripheral nervous system, e.g., cognitive and neurodegenerative disorders, Examples of neurological disorders include, but are not limited to, autonomic function disorders such as hypertension and sleep disorders, and neuropsychiatric disorders, such as depression, schizophrenia, schizoaffective disorder, Korsakoff's psychosis, alcoholism, anxiety disorders, or phobic disorders; learning or memory disorders, e.g., amnesia or age-related memory loss, attention deficit disorder, dysthymic disorder, major depressive disorder, mania, obsessive-compulsive disorder, psychoactive substance use disorders, anxiety, phobias, panic disorder, as well as bipolar affective disorder, e.g., severe bipolar affective (mood) disorder (BP-1), and bipolar affective neurological disorders, e.g., migraine and obesity. Such neurological disorders include, for example, disorders involving neurons, and disorders involving glia, such as astrocytes, oligodendrocytes, ependymal cells, and microglia; cerebral edema, raised intracranial pressure and herniation, and hydrocephalus; malformations and developmental diseases, such as neural tube defects, forebrain anomalies, posterior fossa anomalies, and syringomyelia and hydromyelia; perinatal brain injury; cerebrovascular diseases, such as those related to hypoxia, ischemia, and infarction, including hypotension, hypoperfusion, and low-flow states—global cerebral ischemia and focal cerebral ischemia—infarction from obstruction of local blood supply, intracranial hemorrhage, including intracerebral (intraparenchymal) hemorrhage, subarachnoid hemorrhage and ruptured berry aneurysms, and vascular malformations, hypertensive cerebrovascular disease, including lacunar infarcts, slit hemorrhages, and hypertensive encephalopathy; infections, such as acute meningitis, including acute pyogenic (bacterial) meningitis and acute aseptic (viral) meningitis, acute focal suppurative infections, including brain abscess, subdural empyema, and extradural abscess, chronic bacterial meningoencephalitis, including tuberculosis and mycobacterioses, neurosyphilis, and neuroborreliosis (Lyme disease), viral meningoencephalitis, including arthropod-borne (Arbo) viral encephalitis, Herpes simplex virus Type 1, Herpes simplex virus Type 2, Varicella-zoster virus (Herpes zoster), cytomegalovirus, poliomyelitis, rabies, and human immunodeficiency virus 1, including HIV-1 meningoencephalitis (subacute encephalitis), vacuolar myelopathy, AIDS-associated myopathy, peripheral neuropathy, and AIDS in children, progressive multifocal leukoencephalopathy, subacute sclerosing panencephalitis, fungal meningoencephalitis, other infectious diseases of the nervous system; transmissible spongiform encephalopathies (prion diseases); demyelinating diseases, including multiple sclerosis, multiple sclerosis variants, acute disseminated encephalomyelitis and acute necrotizing hemorrhagic encephalomyelitis, and other diseases with demyelination; degenerative diseases, such as degenerative diseases affecting the cerebral cortex, including Alzheimer's disease and Pick's disease, degenerative diseases of basal ganglia and brain stem, including Parkinsonism, idiopathic Parkinson's disease (paralysis agitans) and other Lewy diffuse body diseases, progressive supranuclear palsy, corticobasal degenration, multiple system atrophy, including striatonigral degenration, Shy-Drager syndrome, and olivopontocerebellar atrophy, and Huntington's disease, senile dementia, Gilles de la Tourette's syndrome, epilepsy, and Jakob-Creutzfieldt disease; spinocerebellar degenerations, including spinocerebellar ataxias, including Friedreich ataxia, and ataxia-telanglectasia, degenerative diseases affecting motor neurons, including amyotrophic lateral sclerosis (motor neuron disease), bulbospinal atrophy (Kennedy syndrome), and spinal muscular atrophy; inborn errors of metabolism, such as leukodystrophies, including Krabbe disease, metachromatic leukodystrophy, adrenoleukodystrophy, Pelizaeus-Merzbacher disease, and Canavan disease, mitochondrial encephalomyopathies, including Leigh disease and other mitochondrial encephalomyopathies; toxic and acquired metabolic diseases, including vitamin deficiencies such as thiamine (vitamin B₁) deficiency and vitamin B₁₂deficiency, neurologic sequelae of metabolic disturbances, including hypoglycemia, hyperglycemia, and hepatic encephatopathy, toxic disorders, including carbon monoxide, methanol, ethanol, and radiation, including combined methotrexate and radiation-induced injury; tumors, such as gliomas, including astrocytoma, including fibrillary (diffuse) astrocytoma and glioblastoma multiforme, pilocytic astrocytoma, pleomorphic xanthoastrocytoma, and brain stem glioma, oligodendroglioma, and ependymoma and related paraventricular mass lesions, neuronal tumors, poorly differentiated neoplasms, including medulloblastoma, other parenchymal tumors, including primary brain lymphoma, germ cell tumors, and pineal parenchymal tumors, meningiomas, metastatic tumors, paraneoplastic syndromes, peripheral nerve sheath tumors, including schwannoma, neurofibroma, and malignant peripheral nerve sheath tumor (malignant schwannoma), and neurocutaneous syndromes (phakomatoses), including neurofibromotosis, including Type 1 neurofibromatosis (NF1) and TYPE 2 neurofibromatosis (NF2), tuberous sclerosis, and Von Hippel-Lindau disease. Further CNS-related disorders include, for example, those listed in the American Psychiatric Association's Diagnostic and Statistical manual of Mental Disorders (DSM), the most current version of which is incorporated herein by reference in its entirety.

As used herein, diseases of the skin (dermal disorders), include but are not limited to, disorders of pigmentation and melanocytes, including but not limited to, vitiligo, freckle, melasma, lentigo, nevocellular nevus, dysplastic nevi, and malignant melanoma; benign epithelial tumors, including but not limited to, seborrheic keratoses, acanthosis nigricans, fibroepithelial polyp, epithelial cyst, keratoacanthoma, and adnexal (appendage) tumors; premalignant and malignant epidermal tumors, including but not limited to, actinic keratosis, squamous cell carcinoma, basal cell carcinoma, and merkel cell carcinoma; tumors of the dermis, including but not limited to, benign fibrous histiocytoma, dermatofibrosarcoma protuberans, xanthomas, and dermal vascular tumors; tumors of cellular immigrants to the skin, including but not limited to, histiocytosis X, mycosis fungoides (cutaneous T-cell lymphoma), and mastocytosis; disorders of epidermal maturation, including but not limited to, ichthyosis; acute inflammatory dermatoses, including but not limited to, urticaria, acute eczematous dermatitis, and erythema multiforme; chronic inflammatory dermatoses, including but not limited to, psoriasis, lichen planus, and lupus erythematosus; blistering (bullous) diseases, including but not limited to, pemphigus, bullous pemphigoid, dermatitis herpetiformis, and noninflammatory blistering diseases: epidermolysis bullosa and porphyria; disorders of epidermal appendages, including but not limited to, acne vulgaris; panniculitis, including but not limited to, erythema nodosum and erythema induratum; and infection and infestation, such as verrucae, molluscum contagiosum, impetigo, superficial fungal infections, and arthropod bites, stings, and infestations.

Additionally, molecules of the invention can play an important role in the regulation of metabolism or pain disorders. Diseases of metabolic imbalance include, but are not limited to, obesity, anorexia nervosa, cachexia, lipid disorders, and diabetes. Examples of pain disorders include, but are not limited to, pain response elicited during various forms of tissue injury, e.g., inflammation, infection, and ischemia, usually referred to as hyperalgesia (described in, for example, Fields (1987) Pain, New York:McGraw-Hill); pain associated with musculoskeletal disorders, e.g., joint pain; tooth pain; headaches; pain associated with surgery; pain related to irritable bowel syndrome; or chest pain.

As used herein, the term “erythroid associated disorders” include disorders involving aberrant (increased or deficient) erythroblast proliferation, e.g., an erythroleukemia, and aberrant (increased or deficient) erythroblast differentiation, e.g., an anemia. Erythrocyte-associated disorders include anemias such as, for example, drug-(chemotherapy-) induced anemias, hemolytic anemias due to hereditary cell membrane abnormalities, such as hereditary spherocytosis, hereditary elliptocytosis, and hereditary pyropoikilocytosis; hemolytic anemias due to acquired cell membrane defects, such as paroxysmal nocturnal hemoglobinuria and spur cell anemia; hemolytic anemias caused by antibody reactions, for example to the RBC antigens, or antigens of the ABO system, Lewis system, Ii system, Rh system, Kidd system, Duffy system, and Kell system; methemoglobinemia; a failure of erythropoiesis, for example, as a result of aplastic anemia, pure red cell aplasia, myelodysplastic syndromes, sideroblastic anemias, and congenital dyserythropoietic anemia; secondary anemia in non-hematolic disorders, for example, as a result of chemotherapy, alcoholism, or liver disease; anemia of chronic disease, such as chronic renal failure; and endocrine deficiency diseases. Another example of an erythroid-associated disorder is erythrocytosis. Erythrocytosis, a disorder of red blood cell overproduction caused by excessive and/or ectopic erythropoietin production, can be caused by cancers, e.g., a renal cell cancer, a hepatocarcinoma, and a central nervous system cancer. Diseases associated with erythrocytosis include polycythemias, e.g., polycythemia vera, secondary polycythemia, and relative polycythemia.

Compounds herein can be used to treat anemias, in particular, anemias associated with cancer chemotherapy, chronic renal failure, malignancies, adult and juvenile rheumatoid arthritis, disorders of haemoglobin synthesis, prematurity, and zidovudine treatment of HIV infection.

Various aspects of the invention are described in further detail below.

Isolated Nucleic Acid Molecules

In one aspect, the invention provides, an isolated or purified, nucleic acid molecule that encodes a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 polypeptide described herein, e.g., a full length 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein or a fragment thereof, e.g., a biologically active portion of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein. Also included is a nucleic acid fragment suitable for use as a hybridization probe, which can be used, e.g., to identify a nucleic acid molecule encoding a polypeptide of the invention, 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 mRNA, and fragments suitable for use as primers, e.g., PCR primers for the amplification or mutation of nucleic acid molecules.

In one embodiment, an isolated nucleic acid molecule of the invention includes the nucleotide sequence shown in SEQ ID NO:1, 3, 4, 6, 7, 9, 10, 12, 13, 15, 16, 18, 53, 55, 61, 63, 67, 69, 78, 80, 88, 90, 100, 102, 113, 115, 122, 124, 129 or 131, or a portion of any of this nucleotide sequence. In one embodiment, the nucleic acid molecule includes sequences encoding the human 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein (i.e., “the coding region” of SEQ ID NO:1, 4, 7, 10, 13, 16, 53, 61, 67, 78, 88, 100, 113, 122 or 129, as shown in SEQ ID NO:3, 6, 9, 12, 15, 18, 55, 63, 69, 80, 90, 102, 115, 124 or 131, respectively), as well as 5′ untranslated sequences and 3′ untranslated sequences. Alternatively, the nucleic acid molecule can include only the coding region of SEQ ID NO:1, 4, 7, 10, 13, 16, 53, 61, 67, 78, 88, 100, 113, 122 or 129 (e.g., SEQ ID NO:3, 6, 9, 12, 15, 18, 55, 63, 69, 80, 90, 102, 115, 124 or 131) and, e.g., no flanking sequences which normally accompany the subject sequence. In another embodiment, the nucleic acid molecule encodes a sequence, corresponding to a fragment of the protein corresponding to domains within SEQ ID NO:2, 5, 8, 11, 14, 17, 54, 62, 68, 79, 89, 101, 114, 123 or 130.

In another embodiment, an isolated nucleic acid molecule of the invention includes a nucleic acid molecule which is a complement of the nucleotide sequence shown in SEQ ID NO:1, 3, 4, 6, 7, 9, 10, 12, 13, 15, 16, 18, 53, 55, 61, 63, 67, 69, 78, 80, 88, 90, 100, 102, 113, 115, 122, 124, 129 or 131, or a portion of any of these nucleotide sequences. In other embodiments, the nucleic acid molecule of the invention is sufficiently complementary to the nucleotide sequence shown in SEQ ID NO:1, 3, 4, 6, 7, 9, 10, 12, 13, 15, 16, 18, 53, 55, 61, 63, 67, 69, 78, 80, 88, 90, 100, 102, 113, 115, 122, 124, 129 or 131 such that it can hybridize to the nucleotide sequence shown in SEQ ID NO:1, 3, 4, 6, 7, 9, 10, 12,13, 15, 16, 18, 53, 55, 61, 63, 67, 69, 78, 80, 88, 90, 100, 102, 113, 115, 122, 124, 129 or 131, thereby forming a stable duplex.

In one embodiment, an isolated nucleic acid molecule of the present invention includes a nucleotide sequence which is at least about: 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more homologous to the entire length of the nucleotide sequence shown in SEQ ID NO:1, 3, 4, 6, 7, 9, 10, 12, 13, 15, 16, 18, 53, 55, 61, 63, 67, 69, 78, 80, 88, 90, 100, 102, 113, 115, 122, 124, 129 or 131, or a portion, preferably of the same length, of any of these nucleotide sequences.

26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 Nucleic Acid Fragments

A nucleic acid molecule of the invention can include only a portion of the nucleic acid sequence of SEQ ID NO:1, 3, 4, 6, 7, 9, 10, 12, 13, 15, 16, 18, 53, 55, 61, 63, 67, 69, 78, 80, 88, 90, 100, 102, 113, 115, 122, 124, 129 or 131. For example, such a nucleic acid molecule can include a fragment which can be used as a probe or primer or a fragment encoding a portion of a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein, e.g., an immunogenic or biologically active portion of a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein. A fragment can comprise those nucleotides of SEQ ID NO:1, 3, 4, 6, 7, 9, 10, 12, 13, 15, 16, 18, 53, 55, 61, 63, 67, 69, 78, 80, 88, 90, 100, 102, 113, 115, 122, 124, 129 or 131, which encode a domain of human 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843. The nucleotide sequence determined from the cloning of the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 gene allows for the generation of probes and primers designed for use in identifying and/or cloning other 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 family members, or fragments thereof, as well as 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 homologs, or fragments thereof, from other species.

In another embodiment, a nucleic acid includes a nucleotide sequence that includes part, or all, of the coding region and extends into either (or both) the 5′ or 3′ noncoding region. Other embodiments include a fragment which includes a nucleotide sequence encoding an amino acid fragment described herein. Nucleic acid fragments can encode a specific domain or site described herein or fragments thereof, particularly fragments thereof which are at least 100 amino acids in length. Fragments also include nucleic acid sequences corresponding to specific amino acid sequences described above or fragments thereof. Nucleic acid fragments should not to be construed as encompassing those fragments that may have been disclosed prior to the invention.

A nucleic acid fragment can include a sequence corresponding to a domain, region, or functional site described herein. A nucleic acid fragment can also include one or more domain, region, or functional site described herein. Thus, for example, a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 nucleic acid fragment can include a sequence corresponding to a domain, as described herein.

26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 probes and primers are provided. Typically a probe/primer is an isolated or purified oligonucleotide. The oligonucleotide typically includes a region of nucleotide sequence that hybridizes under stringent conditions to at least about 7, 12 or 15, preferably about 20 or 25, more preferably about 30, 35, 40, 45, 50, 55, 60, 65, or 75 consecutive nucleotides of a sense or antisense sequence of SEQ ID NO:1, 3, 4, 6, 7, 9, 10, 12, 13, 15, 16, 18, 53, 55, 61, 63, 67, 69, 78, 80, 88, 90, 100, 102, 113, 115, 122, 124, 129 or 131, or of a naturally occurring allelic variant or mutant of SEQ ID NO:1, 3, 4, 6, 7, 9, 10, 12, 13, 15, 16, 18, 53, 55, 61, 63, 67, 69, 78, 80, 88, 90, 100, 102, 113, 115, 122, 124, 129 or 131.

In a preferred embodiment the nucleic acid is a probe which is at least 5 or 10, and less than 200, more preferably less than 100, or less than 50, base pairs in length. It should be identical, or differ by 1, or less than in 5 or 10 bases, from a sequence disclosed herein. If alignment is needed for this comparison the sequences should be aligned for maximum homology. “Looped” out sequences from deletions or insertions, or mismatches, are considered differences.

A probe or primer can be derived from the sense or anti-sense strand of a nucleic acid which encodes a domain identified in the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 sequences.

In another embodiment a set of primers is provided, e.g., primers suitable for use in a PCR, which can be used to amplify a selected region of a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 sequence, e.g., a domain, region, site or other sequence described herein. The primers should be at least 5, 10, or 50 base pairs in length and less than 100, or less than 200, base pairs in length. The primers should be identical, or differ by one base from a sequence disclosed herein or from a naturally occurring variant.

A nucleic acid fragment can encode an epitope bearing region of a polypeptide described herein.

A nucleic acid fragment encoding a “biologically active portion of a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 polypeptide” can be prepared by isolating a portion of the nucleotide sequence of SEQ ID NO:1, 3, 4, 6, 7, 9, 10, 12, 13, 15, 16, 18, 53, 55, 61, 63, 67, 69, 78, 80, 88, 90, 100, 102, 113, 115, 122, 124, 129 or 131, which encodes a polypeptide having a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 biological activity (e.g., the biological activities of the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 proteins are described herein), expressing the encoded portion of the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein (e.g., by recombinant expression in vitro) and assessing the activity of the encoded portion of the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein. A nucleic acid fragment encoding a biologically active portion of a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 polypeptide, can comprise a nucleotide sequence which is greater than 300 or more nucleotides in length.

In preferred embodiments, a nucleic acid includes a nucleotide sequence which is about 300, 400, 500, 600, 700, 800, 900, 1000, 1100, 1200, 1300, 1400, 1500, 1600, 1700, 1800, 1900, 2000, 2100, 2200, 2300, 2400, 2500, 2600, 2700, 2800, 2900, 3000, 3100, 3200, 3300, 3400, 3500, 3600, 3700, 3800, 3900, 4000, 4100, 4200, 4300, 4400, 4500, 4600, 4700, 4800, 4900, 5000, 5100, 5200, 5300 or more nucleotides in length and hybridizes under stringent hybridization conditions to a nucleic acid molecule of SEQ ID NO:1, 3, 4, 6, 7, 9, 10, 12, 13, 15, 16, 18, 53, 55, 61, 63, 67, 69, 78, 80, 88, 90, 100, 102, 113, 115, 122, 124, 129 or 131.

26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 Nucleic Acid Variants

The invention further encompasses nucleic acid molecules that differ from the nucleotide sequence shown in SEQ ID NO:1, 3, 4, 6, 7, 9, 10, 12, 13, 15, 16, 18, 53, 55, 61, 63, 67, 69, 78, 80, 88, 90, 100, 102, 113, 115, 122, 124, 129 or 131. Such differences can be due to degeneracy of the genetic code (and result in a nucleic acid which encodes the same 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 proteins as those encoded by the nucleotide sequence disclosed herein. In another embodiment, an isolated nucleic acid molecule of the invention has a nucleotide sequence encoding a protein having an amino acid sequence which differs, by at least 1, but less than 5, 10, 20, 50, or 100 amino acid residues that shown in SEQ ID NO:2, 5, 8, 11, 14, 17, 54, 62, 68, 79, 89, 101, 114, 123 or 130. If alignment is needed for this comparison the sequences should be aligned for maximum homology. “Looped” out sequences from deletions or insertions, or mismatches, are considered differences.

Nucleic acids of the inventor can be chosen for having codons, which are preferred, or non-preferred, for a particular expression system. E.g., the nucleic acid can be one in which at least one codon, at preferably at least 10%, or 20% of the codons has been altered such that the sequence is optimized for expression in E. coli, yeast, human, insect, or CHO cells.

Nucleic acid variants can be naturally occurring, such as allelic variants (same locus), homologs (different locus), and orthologs (different organism) or can be non naturally occurring. Non-naturally occurring variants can be made by mutagenesis techniques, including those applied to polynucleotides, cells, or organisms. The variants can contain nucleotide substitutions, deletions, inversions and insertions. Variation can occur in either or both the coding and non-coding regions. The variations can produce both conservative and non-conservative amino acid substitutions (as compared in the encoded product).

In a preferred embodiment, the nucleic acid differs from that of SEQ ID NO:1, 3, 4, 6, 7, 9, 10, 12, 13, 15, 16, 18, 53, 55, 61, 63, 67, 69, 78, 80, 88, 90, 100, 102, 113, 115, 122, 124, 129 or 131, e.g., as follows: by at least one but less than 10, 20, 30, or 40 nucleotides; at least one but less than 1%, 5%, 10% or 20% of the nucleotides in the subject nucleic acid. If necessary for this analysis the sequences should be aligned for maximum homology. “Looped” out sequences from deletions or insertions, or mismatches, are considered differences.

Orthologs, homologs, and allelic variants can be identified using methods known in the art. These variants comprise a nucleotide sequence encoding a polypeptide that is 50%, at least about 55%, typically at least about 70-75%, more typically at least about 80-85%, and most typically at least about 90-95% or more identical to the nucleotide sequence shown in SEQ ID NO:2, 5, 8, 11, 14, 17, 54, 62, 68, 79, 89, 101, 114, 123 or 130 or a fragment of this sequence. Such nucleic acid molecules can readily be identified as being able to hybridize under stringent conditions, to the nucleotide sequence shown in SEQ ID NO:2, 5, 8, 11, 14, 17, 54, 62, 68, 79, 89, 101, 114, 123 or 130 or a fragment of the sequence. Nucleic acid molecules corresponding to orthologs, homologs, and allelic variants of the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 cDNAs of the invention can further be isolated by mapping to the same chromosome or locus as the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 gene.

Preferred variants include those that are correlated with activities specific to the molecules of the invention, i.e. arginine methyltransferase activity, glycosyltransferase activity, gamma-glutamyltraspeptidase activity, phosphoribosylglycinamide transferase activity, acyltransferase activity, acyl-CoA dehydrogenase activity, fatty acid amide hydrolase activity, aminotransferase activity, zinc carboxypeptidase activity, protein kinase activity, DEAD helicase activity, short-chain dehydrogenase/reductase activity or phosphatase activity, or other.

Allelic variants of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843, e.g., human 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843, include both functional and non-functional proteins. Functional allelic variants are naturally occurring amino acid sequence variants of the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein within a population that maintain the ability to bind a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 ligand or substrate and/or modulate cell proliferation and/or migration mechanisms. Functional allelic variants will typically contain only conservative substitution of one or more amino acids of SEQ ID NO:2, 5, 8, 11, 14, 17, 54, 62, 68, 79, 89, 101, 114, 123 or 130, or substitution, deletion or insertion of non-critical residues in non-critical regions of the protein. Non-functional allelic variants are naturally-occurring amino acid sequence variants of the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843, e.g., human 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843, protein within a population that do not have the ability to bind a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 ligand or substrate and/or modulate cell proliferation and/or migration mechanisms. Non-functional allelic variants will typically contain a non-coservative substitution, a deletion, or insertion, or premature truncation of the amino acid sequence of SEQ ID NO:2, 5, 8, 11, 14, 17, 54, 62, 68, 79, 89, 101, 114, 123 or 130, or a substitution, insertion, or deletion in critical residues or critical regions of the protein.

Moreover, nucleic acid molecules encoding other 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 family members and, thus, which have a nucleotide sequence which differs from the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 sequences of SEQ ID NO:1, 3, 4, 6, 7, 9, 10, 12, 13, 15, 16, 18, 53, 55, 61, 63, 67, 69, 78, 80, 88, 90, 100, 102, 113, 115, 122, 124, 129 or 131 are intended to be within the scope of the invention.

Antisense Nucleic Acid Molecules, Ribozymes and Modified 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 Nucleic Acid Molecules

In another aspect, the invention features, an isolated nucleic acid molecule which is antisense to 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843. An “antisense” nucleic acid can include a nucleotide sequence which is complementary to a “sense” nucleic acid encoding a protein, e.g., complementary to the coding strand of a double-stranded cDNA molecule or complementary to an mRNA sequence. The antisense nucleic acid can be complementary to an entire 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 coding strand, or to only a portion thereof (e.g., the coding region of human 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 corresponding to SEQ ID NO:3, 6, 9, 12, 15, 18, 55, 63, 69, 80, 90, 102, 115, 124 or 131, respectively). In another embodiment, the antisense nucleic acid molecule is antisense to a “noncoding region” of the coding strand of a nucleotide sequence encoding 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 (e.g., the 5′ and 3′ untranslated regions).

An antisense nucleic acid can be designed such that it is complementary to the entire coding region of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 mRNA, but more preferably is an oligonucleotide which is antisense to only a portion of the coding or noncoding region of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 mRNA. For example, the antisense oligonucleotide can be complementary to the region surrounding the translation start site of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 mRNA, e.g., between the −10 and +10 regions of the target gene nucleotide sequence of interest. An antisense oligonucleotide can be, for example, about 7, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, or more nucleotides in length.

An antisense nucleic acid of the invention can be constructed using chemical synthesis and enzymatic ligation reactions using procedures known in the art. For example, an anfisense nucleic acid (e.g., an antisense oligonucleotide) can be chemically synthesized using naturally occurring nucleotides or variously modified nucleotides designed to increase the biological stability of the molecules or to increase the physical stability of the duplex formed between the antisense and sense nucleic acids, e.g., phosphorothioate derivatives and acridine substituted nucleotides can be used. The antisense nucleic acid also can be produced biologically using an expression vector into which a nucleic acid has been subcloned in an antisense orientation (i.e., RNA transcribed from the inserted nucleic acid will be of an antisense orientation to a target nucleic acid of interest, described further in the following subsection).

The antisense nucleic acid molecules of the invention are typically administered to a subject (e.g., by direct injection at a tissue site), or generated in situ such that they hybridize with or bind to cellular mRNA and/or genomic DNA encoding a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein to thereby inhibit expression of the protein, e.g., by inhibiting transcription and/or translation. Alternatively, antisense nucleic acid molecules can be modified to target selected cells and then administered systemically. For systemic administration, antisense molecules can be modified such that they specifically or selectively bind to receptors or antigens expressed on a selected cell surface, e.g., by linking the antisense nucleic acid molecules to peptides or antibodies which bind to cell surface receptors or antigens. The antisense nucleic acid molecules can also be delivered to cells using the vectors described herein. To achieve sufficient intracellular concentrations of the antisense molecules, vector constructs in which the antisense nucleic acid molecule is placed under the control of a strong pol II or pol III promoter are preferred.

In yet another embodiment, the antisense nucleic acid molecule of the invention is an α-anomeric nucleic acid molecule. An α-anomeric nucleic acid molecule forms specific double-stranded hybrids with complementary RNA in which, contrary to the usual β-units, the strands run parallel to each other (Gaultier et al. (1987) Nucleic Acids. Res. 15:6625-6641). The antisense nucleic acid molecule can also comprise a 2′-o-methylribonucleotide (Inoue et al. (1987) Nucleic Acids Res. 15:6131-6148) or a chimeric RNA-DNA analogue (Inoue et al. (1987) FEBS Lett. 215:327-330).

In still another embodiment, an antisense nucleic acid of the invention is a ribozyme. A ribozyme having specificity for a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843-encoding nucleic acid can include one or more sequences complementary to the nucleotide sequence of a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 cDNA disclosed herein (i.e., SEQ ID NO:1, 3, 4, 6, 7, 9, 10, 12, 13, 15, 16, 18, 53, 55, 61, 63, 67, 69, 78, 80, 88, 90, 100, 102, 113, 115, 122, 124, 129 or 131), and a sequence having known catalytic sequence responsible for mRNA cleavage (see U.S. Pat. No. 5,093,246 or Haselhoff and Gerlach (1988) Nature 334:585-591). For example, a derivative of a Tetrahymena L-19 IVS RNA can be constructed in which the nucleotide sequence of the active site is complementary to the nucleotide sequence to be cleaved in a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843-encoding mRNA. See, e.g., Cech et al. U.S. Pat. No. 4,987,071; and Cech et al. U.S. Pat. No. 5,116,742. Alternatively, 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 mRNA can be used to select a catalytic RNA having a specific ribonuclease activity from a pool of RNA molecules. See, e.g., Bartel and Szostak (1993) Science 261:1411-1418.

26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 gene expression can be inhibited by targeting nucleotide sequences complementary to the regulatory region of the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 (e.g., the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 promoter and/or enhancers) to form triple helical structures that prevent transcription of the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 gene in target cells. See generally, Helene (1991) Anticancer Drug Des. 6:569-84; Helene (1992) Ann. N.Y Acad. Sci. 660:27-36; and Maher (1992) Bioassays 14:807-15. The potential sequences that can be targeted for triple helix formation can be increased by creating a so-called “switchback” nucleic acid molecule. Switchback molecules are synthesized in an alternating 5′-3′, 3′-5′ manner, such that they base pair with first one strand of a duplex and then the other, eliminating the necessity for a sizeable stretch of either purines or pyrimidines to be present on one strand of a duplex.

The invention also provides detectably labeled oligonucleotide primer and probe molecules. Typically, such labels are chemiluminescent, fluorescent, radioactive, or calorimetric.

A 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 nucleic acid molecule can be modified at the base moiety, sugar moiety or phosphate backbone to improve, e.g., the stability, hybridization, or solubility of the molecule. For example, the deoxyribose phosphate backbone of the nucleic acid molecules can be modified to generate peptide nucleic acids (see Hyrup et al. (1996) Bioorganic & Medicinal Chemistry 4: 5-23).

As used herein, the terms “peptide nucleic acid” or “PNA” refers to a nucleic acid mimic, e.g., a DNA mimic, in which the deoxyribose phosphate backbone is replaced by a pseudopeptide backbone and only the four natural nucleobases are retained. The neutral backbone of a PNA can allow for specific hybridization to DNA and RNA under conditions of low ionic strength. The synthesis of PNA oligomers can be performed using standard solid phase peptide synthesis protocols as described in Hyrup et al. (1996) supra; Perry-O'Keefe et al. (1996) Proc. Natl. Acad. Sci. 93: 14670-675.

PNAs of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 nucleic acid molecules can be used in therapeutic and diagnostic applications. For example, PNAs can be used as antisense or antigene agents for sequence-specific modulation of gene expression by, for example, inducing transcription or translation arrest or inhibiting replication. PNAs of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 nucleic acid molecules can also be used in the analysis of single base pair mutations in a gene, (e.g., by PNA-directed PCR clamping); as ‘artificial restriction enzymes’ when used in combination with other enzymes, (e.g., SI nucleases (Hyrup et al. (1996) supra)); or as probes or primers for DNA sequencing or hybridization (Hyrup et al. (1996) supra; Perry-O'Keefe supra).

In other embodiments, the oligonucleotide can include other appended groups such as peptides (e.g., for targeting host cell receptors in vivo), or agents facilitating transport across the cell membrane (see, e.g., Letsinger et al. (1989) Proc. Natl. Acad. Sci. USA 86:6553-6556; Lemaitre et al. (1987) Proc. Natl. Acad. Sci. USA 84:648-652; PCT Publication No. WO88/09810) or the blood-brain barrier (see, e.g., PCT Publication No. WO89/10134). In addition, oligonucleotides can be modified with hybridization-triggered cleavage agents (see, e.g., Krol et al. (1988) Bio-Techniques 6:958-976) or intercalating agents. (see, e.g., Zon (1988) Pharm. Res. 5:539-549). To this end, the oligonucleotide can be conjugated to another molecule, (e.g., a peptide, hybridization triggered cross-linking agent, transport agent, or hybridization-triggered cleavage agent).

The invention also includes molecular beacon oligonucleotide primer and probe molecules having at least one region which is complementary to a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 nucleic acid of the invention, two complementary regions one having a fluorophore and one a quencher such that the molecular beacon is useful for quantitating the presence of the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 nucleic acid of the invention in a sample. Molecular beacon nucleic acids are described, for example, in Lizardi et al., U.S. Pat. No. 5,854,033; Nazarenko et al., U.S. Pat. No. 5,866,336, and Livak et al., U.S. Pat. 5,876,930.

Isolated 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 Polypeptides

In another aspect, the invention features, an isolated 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein, or fragment, e.g., a biologically active portion, for use as immunogens or antigens to raise or test (or more generally to bind) anti-26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 antibodies. 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein can be isolated from cells or tissue sources using standard protein purification techniques. 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein or fragments thereof can be produced by recombinant DNA techniques or synthesized chemically.

Polypeptides of the invention include those which arise as a result of the existence of multiple genes, alternative transcription events, alternative RNA splicing events, and alternative translational and post-translational events. The polypeptide can be expressed in systems, e.g., cultured cells, which result in substantially the same post-translational modifications present when the polypeptide is expressed in a native cell, or in systems which result in the alteration or omission of post-translational modifications, e.g., glycosylation or cleavage, present in a native cell.

In a preferred embodiment, a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 polypeptide has one or more of the following characteristics: it has the ability: (i) to transfer an activated sugar residue to an acceptor molecule; (ii) to modulate the processing, folding, and secretion of proteins; (iii) to transport amino acids in the form of their gamma-glutamyl derivatives; (iv) to regulate the metabolism of glutathione; (v) to regulate the synthesis of purines; (vi) to modulate cell division and proliferation; (vii) to modulate cell death; (viii) to transfer an acyl chain to a lipid precursor; (ix) to regulate lipid biosynthesis; (x) to catalyze the transfer of hydrogen and electrons from one compound to another; (xi) to catalyze the 1,θ-dehydrogenation of fatty acyl-CoA derivatives; (xii) to bind and catabolize fatty acid amides; (xiii) to modulate metabolism, e.g., amino acid metabolism; (xiv) to bind an amino acid, e.g., L-alanine; (xv) to bind an oxo acid, e.g., pyruvate; (xvi) to modulate the formation of a zinc ion complex with a carbonyl group of a substrate polypeptide and polarization of the carbon-oxygen bond; (xvii) to modulate formation of a tetrahedral intermediate due to attack of the carbonyl carbon by water in a reaction assisted by a carboxylate side chain of glutamate; (xviii) to modulate the production of a dianion intermediate by rapid ionization of the tetrahedral intermediate produced; (xix) to modulate ATP dependent nucleic acid unwinding; (xx) to modulate RNA metabolism (e.g., nuclear transcription, and mRNA splicing); (xxi) to modulate steroid biosynthesis or metabolism (breakdown); (xxii) to catalyze the removal of a phosphate group attached to a tyrosine residue in a protein; (xxiii) to catalyze the removal of a phosphate group attached to a serine or threonine residue in a protein; (xxiv) to modulate an intracellular signaling pathway, e.g., a MAP kinase or ERK kinase pathway; (xxv) to regulate the transmission of signals from cellular receptors, e.g., cardiac cell growth factor receptors; (xxvi) it has a molecular weight, e.g., a deduced molecular weight, preferably ignoring any contribution of post translational modifications, amino acid composition or other physical characteristic of a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 polypeptide, e.g., a polypeptide of SEQ ID NO:2, 5, 8, 11, 14, 17, 54, 62, 68, 79, 89, 101, 114, 123 or 130; (xxvii) it has an overall sequence similarity of at least 60%, preferably at least 70%, more preferably at least 80, 90, or 95%, with a polypeptide of SEQ ID NO:2, 5, 8, 11, 14, 17, 54, 62, 68, 79, 89, 101, 114, 123 or 130; (xxviii) it is expressed in a multitude of human tissues and cell lines (refer to section for each molecule of the invention); and (xxix) it has specific domains which are preferably about 70%, 80%, 90% or 95% identical to the identified amino acid residues of SEQ ID NO:2, 5, 8, 11, 14, 17, 54, 62, 68, 79, 89, 101, 114, 123 or 130 (refer to section for each molecule of the invention for domain names and locations within amino acid sequence).

In a preferred embodiment the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein, or fragment thereof, differs from the corresponding sequence in SEQ ID NO:2, 5, 8, 11, 14, 17, 54, 62, 68, 79, 89, 101, 114, 123 or 130. In one embodiment it differs by at least one but by less than 15, 10 or 5 amino acid residues. In another it differs from the corresponding sequence in SEQ ID NO:2, 5, 8, 11, 14, 17, 54, 62, 68, 79, 89, 101, 114, 123 or 130 by at least one residue but less than 20%, 15%, 10% or 5% of the residues in it differ from the corresponding sequence in SEQ ID NO:2, 5, 8, 11, 14, 17, 54, 62, 68, 79, 89, 101, 114, 123 or 130. (If this comparison requires alignment the sequences should be aligned for maximum homology. “Looped” out sequences from deletions or insertions, or mismatches, are considered differences.) The differences are, preferably, differences or changes at a non-essential residue or a conservative substitution. In a preferred embodiment the differences are not in the identified or conserved domain(s) within SEQ ID NO:2, 5, 8, 11, 14, 17, 54, 62, 68, 79, 89, 101, 114, 123 or 130. In another embodiment one or more differences are in the cidentified or conserved domain(s) within SEQ ID NO:2, 5, 8, 11, 14, 17, 54, 62, 68, 79, 89, 101, 114, 123 or 130.

Other embodiments include a protein that contains one or more changes in amino acid sequence, e.g., a change in an amino acid residue which is not essential for activity. Such 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 proteins differ in amino acid sequence from SEQ ID NO:2, 5, 8, 11, 14, 17, 54, 62, 68, 79, 89, 101, 114, 123 or 130, yet retain biological activity.

In one embodiment, the protein includes an amino acid sequence at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98% or more homologous to SEQ ID NO:2, 5, 8, 11, 14, 17, 54, 62, 68, 79, 89, 101, 114, 123 or 130.

A 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein or fragment is provided which varies from the sequence of SEQ ID NO:2, 5, 8, 11, 14, 17, 54, 62, 68, 79, 89, 101, 114, 123 or 130 in regions defined by amino acids that are not within identified or conserved domains or regions by at least one but by less than 15, 10 or 5 amino acid residues in the protein or fragment but which does not differ from SEQ ID NO:2, 5, 8, 11, 14, 17, 54, 62, 68, 79, 89, 101, 114, 123 or 130 in regions defined by amino acids that are within identified or conserved domains or regions. (If this comparison requires alignment the sequences should be aligned for maximum homology. “Looped” out sequences from deletions or insertions, or mismatches, are considered differences.) In some embodiments the difference is at a non-essential residue or is a conservative substitution, while in others the difference is at an essential residue or is a non-conservative substitution.

In one embodiment, a biologically active portion of a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein includes an identified domain (refer to section for each molecule of the invention). Moreover, other biologically active portions, in which other regions of the protein are deleted, can be prepared by recombinant techniques and evaluated for one or more of the functional activities of a native 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein.

In a preferred embodiment, the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein has an amino acid sequence shown in SEQ ID NO:2, 5, 8, 11, 14, 17, 54, 62, 68, 79, 89, 101, 114, 123 or 130. In other embodiments, the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein is sufficiently or substantially identical to SEQ ID NO:2, 5, 8, 11, 14, 17, 54, 62, 68, 79, 89, 101, 114, 123 or 130. In yet another embodiment, the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein is sufficiently or substantially identical to. SEQ ID NO:2, 5, 8, 11, 14, 17, 54, 62, 68, 79, 89, 101, 114, 123 or 130 and retains the functional activity of the protein of SEQ ID NO:2, 5, 8, 11, 14, 17, 54, 62, 68, 79, 89, 101, 114, 123 or 130, as described in detail in the subsections above.

26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 Chimeric or Fusion Proteins

In another aspect, the invention provides 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 chimeric or fusion proteins. As used herein, a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 “chimeric protein” or “fusion protein” includes a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 polypeptide linked to a non-26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216,25206 or 8843 polypeptide. A “non-26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 polypeptide” refers to a polypeptide having an amino acid sequence corresponding to a protein which is not substantially homologous to the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein, e.g., a protein which is different from the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein and which is derived from the same or a different organism. The 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 polypeptide of the fusion protein can correspond to all or a portion e.g., a fragment described herein of a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 amino acid sequence. In a preferred embodiment, a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 fusion protein includes at least one (or two) biologically active portion of a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein. The non-26199, 33530, 33949, 47148, 50226, 58764,62113, 32144,32235,23565, 13305, 14911, 86216, 25206 or 8843 polypeptide can be fused to the N-terminus or C-terminus of the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 polypeptide.

The fusion protein can include a moiety which has a high affinity for a ligand. For example, the fusion protein can be a GST-26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 fusion protein in which the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 sequences are fused to the C-terminus of the GST sequences. Such fusion proteins can facilitate the purification of recombinant 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843. Alternatively, the fusion protein can be a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein containing a heterologous signal sequence at its N-terminus. In certain host cells (e.g., mammalian host cells), expression and/or secretion of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 can be increased through use of a heterologous signal sequence.

Fusion proteins can include all or a part of a serum protein, e.g., a portion of an immunoglobulin (e.g., IgG, IgA, or IgE), e.g., an Fc region and/or the hinge C1 and C2 sequences of an immunoglobulin or human serum albumin.

The 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 fusion proteins of the invention can be incorporated into pharmaceutical compositions and administered to a subject in vivo. The 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 fusion proteins can be used to affect the bioavailability of a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 substrate. 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 fusion proteins can be useful therapeutically for the treatment of disorders caused by, for example, (i) aberrant modification or mutation of a gene encoding a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein; (ii) mis-regulation of the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 gene; and (iii) aberrant post-translational modification of a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein.

Moreover, the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843-fusion proteins of the invention can be used as immunogens to produce anti-26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 antibodies in a subject, to purify 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 ligands and in screening assays to identify molecules which inhibit the interaction of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 with a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 substrate.

Expression vectors are commercially available that already encode a fusion moiety (e.g., a GST polypeptide). A 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843-encoding nucleic acid can be cloned into such an expression vector such that the fusion moiety is linked in-frame to the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein.

Variants of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 Proteins

In another aspect, the invention also features a variant of a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 polypeptide, e.g., which functions as an agonist (mimetics) or as an antagonist. Variants of the.26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 proteins can be generated by mutagenesis, e.g., discrete point mutation, the insertion or deletion of sequences or the truncation of a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein. An agonist of the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 proteins can retain substantially the same, or a subset, of the biological activities of the naturally occurring form of a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein. An antagonist of a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein can inhibit one or more of the activities of the naturally occurring form of the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein by, for example, competitively modulating a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843-mediated activity of a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein. Thus, specific biological effects can be elicited by treatment with a variant of limited function. Preferably, treatment of a subject with a variant having a subset of the biological activities of the naturally occurring form of the protein has fewer side effects in a subject relative to treatment with the naturally occurring form of the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein.

Variants of a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein can be identified by screening combinatorial libraries of mutants, e.g., truncation mutants, of a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein for agonist or antagonist activity.

Libraries of fragments e.g., N terminal, C terminal, or internal fragments, of a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein coding sequence can be used to generate a variegated population of fragments for screening and subsequent selection of variants of a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein.

Variants in which a cysteine residues is added or deleted or in which a residue which is glycosylated is added or deleted are particularly preferred.

Methods for screening gene products of combinatorial libraries made by point mutations or truncation, and for screening cDNA libraries for gene products having a selected property are known in the art. Recursive ensemble mutagenesis (REM), a new technique which enhances the frequency of functional mutants in the libraries, can be used in combination with the screening assays to identify 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 variants (Arkin and Yourvan (1992) Proc. Natl. Acad. Sci. USA 89:7811-7815; Delgrave et al. (1993) Protein Engineering 6:327-331).

Cell based assays can be exploited to analyze a variegated 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 library. For example, a library of expression vectors can be transfected into a cell line, e.g., a cell line, which ordinarily responds to 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 in a substrate-dependent manner. The transfected cells are then contacted with 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 and the effect of the expression of the mutant on signaling by the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 substrate can be detected, e.g., by measuring either arginine methyltransferase activity, glycosyltransferase activity, gamma-glutamyltraspeptidase activity, phosphoribosylglycinamide transferase activity, acyltransferase activity, acyl-CoA dehydrogenase activity, fatty acid amide hydrolase activity, aminotransferase activity, zinc carboxypeptidase activity, protein kinase activity, DEAD helicase activity, short-chain dehydrogenase/reductase activity or phosphatase activity, or other activity. Plasmid DNA can then be recovered from the cells which score for inhibition, or alternatively, potentiation of signaling by the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 substrate, and the individual clones further characterized.

In another aspect, the invention features a method of making a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 polypeptide, e.g., a peptide having a non-wild type activity, e.g., an antagonist, agonist, or super agonist of a naturally occurring 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 polypeptide, e.g., a naturally occurring 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 polypeptide. The method includes altering the sequence of a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 polypeptide, e.g., altering the sequence, e.g., by substitution or deletion of one or more residues of a non-conserved region, a domain or residue disclosed herein, and testing the altered polypeptide for the desired activity.

In another aspect, the invention features a method of making a fragment or analog of a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 polypeptide a biological activity of a naturally occurring 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 polypeptide. The method includes altering the sequence, e.g., by substitution or deletion of one or more residues, of a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 polypeptide, e.g., altering the sequence of a non-conserved region, or a domain or residue described herein, and testing the altered polypeptide for the desired activity.

Anti-26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 Antibodies

In another aspect, the invention provides an anti-26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 antibody. The term “antibody” as used herein refers to an immunoglobulin molecule or immunologically active portion thereof, i.e., an antigen-binding portion. Examples of immunologically active portions of immunoglobulin molecules include scFV and dcFV fragments, Fab and F(ab′) ₂fragments which can be generated by treating the antibody with an enzyme such as papain or pepsin, respectively.

The antibody can be a polyclonal, monoclonal, recombinant, e.g., a chimeric or humanized, fully human, non-human, e.g., murine, or single chain antibody. In a preferred embodiment it has effector function and can fix complement. The antibody can be coupled to a toxin or imaging agent.

A full-length 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein or, antigenic peptide fragment of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 can be used as an immunogen or can be used to identify anti-26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 antibodies made with other immunogens, e.g., cells, membrane preparations, and the like. The antigenic peptide of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 should include at least 8 amino acid residues of the amino acid sequence shown in SEQ ID NO:2, 5, 8, 11, 14, 17, 54, 62, 68, 79, 89, 101, 114, 123 or 130 and encompasses an epitope of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843. Preferably, the antigenic peptide includes at least 10 amino acid residues, more preferably at least 15 amino acid residues, even more preferably at least 20 amino acid residues, and most preferably at least 30 amino acid residues.

Fragments of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 which include hydrophilic regions of SEQ ID NO:2, 5, 8, 11, 14, 17, 54, 62, 68, 79, 89, 101, 114, 123 or 130 can be used to make, e.g., used as immunogens or used to characterize the specificity of an antibody, antibodies against hydrophilic regions of the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein. Similarly, fragments of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 which include hydrophobic regions of SEQ ID NO:2, 5, 8, 11, 14, 17, 54, 62, 68, 79, 89, 101, 114, 123 or 130 can be used to make an antibody against a hydrophobic region of the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein; fragments of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216,25206 or 8843 which include residues within extra cellular domain(s) of SEQ ID NO:2, 5, 8, 11, 14, 17, 54, 62, 68, 79, 89, 101, 114, 123 or 130 can be used to make an antibody against an extracellular or non-cytoplasmic region of the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein; fragments of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 which include residues within intracellular regions of SEQ ID NO:2, 5, 8, 11, 14, 17, 54, 62, 68, 79, 89, 101, 114, 123 or 130 can be used to make an antibody against an intracellular region of the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein; a fragment of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 which include residues within identified or conserved domains of SEQ ID NO:2, 5, 8, 11, 14, 17, 54, 62, 68, 79, 89, 101, 114, 123 or 130 can be used to make an antibody against the identified or conserved domain of the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein.

Antibodies reactive with, or specific or selective for, any of these regions, or other regions or domains described herein are provided.

Preferred epitopes encompassed by the antigenic peptide are regions of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 located on the surface of the protein, e.g., hydrophilic regions, as well as regions with high antigenicity. For example, an Emini surface probability analysis of the human 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein sequence can be used to indicate the regions that have a particularly high probability of being localized to the surface of the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein and are thus likely to constitute surface residues useful for targeting antibody production.

In a preferred embodiment the antibody can bind to the extracellular portion of the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein, e.g., it can bind to a whole cell which expresses the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein. In another embodiment, the antibody binds an intracellular portion of the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein.

In a preferred embodiment the antibody binds an epitope on any domain or region on 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 proteins described herein.

Additionally, chimeric, humanized, and completely human antibodies are also within the scope of the invention. Chimeric, humanized, but most preferably, completely human antibodies are desirable for applications which include repeated administration, e.g., therapeutic treatment of human patients, and some diagnostic applications.

Chimeric and humanized monoclonal antibodies, comprising both human and non-human portions, can be made using standard recombinant DNA techniques. Such chimeric and humanized monoclonal antibodies can be produced by recombinant DNA techniques known in the art, for example using methods described in Robinson et al. International Application No. PCT/US86/02269; Akira, et al. European Patent Application 184,187; Taniguchi, European Patent Application 171,496; Morrison et al. European Patent Application 173,494; Neuberger et al. PCT International Publication No. WO 86/01533; Cabilly et al. U.S. Pat. No. 4,816,567; Cabilly et al. European Patent Application 125,023; Better et al. (1988) Science 240:1041-1043; Liu et al. (1987) Proc. Natl. Acad. Sci. USA 84:3439-3443; Liu et al. (1987) J. Immunol. 139:3521-3526; Sun et al. (1987) Proc. Natl. Acad. Sci. USA 84:214-218; Nishimura et al. (1987) Canc. Res. 47:999-1005; Wood et al. (1985) Nature 314:446-449; and Shaw et al. (1988) J. Natl. Cancer Inst. 80:1553-1559).

A humanized or complementarity determining region (CDR)-grafted antibody will have at least one or two, but generally all three recipient CDR's (of heavy and or light immuoglobulin chains) replaced with a donor CDR. The antibody may be replaced with at least a portion of a non-human CDR or only some of the CDR's may be replaced with non-human CDR's. It is only necessary to replace the number of CDR's required for binding of the humanized antibody to a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 or a fragment thereof. Preferably, the donor will be a rodent antibody, e.g., a rat or mouse antibody, and the recipient will be a human framework or a human consensus framework. Typically, the immunoglobulin providing the CDR's is called the “donor” and the immunoglobulin providing the framework is called the “acceptor.” In one embodiment, the donor immunoglobulin is a non-human (e.g., rodent). The acceptor framework is a naturally-occuring (e.g., a human) framework or a consensus framework, or a sequence about 85% or higher, preferably 90%, 95%, 99% or higher identical thereto.

As used herein, the term “consensus sequence” refers to the sequence formed from the most frequently occurring amino acids (or nucleotides) in a family of related sequences (See e.g., Winnaker, (1987) From Genes to Clones (Verlagsgesellschaft, Weinheim, Germany). In a family of proteins, each position in the consensus sequence is occupied by the amino acid occurring most frequently at that position in the family. If two amino acids occur equally frequently, either can be included in the consensus sequence. A “consensus framework” refers to the framework region in the consensus immunoglobulin sequence.

An antibody can be humanized by methods known in the art. Humanized antibodies can be generated by replacing sequences of the Fv variable region which are not directly involved in antigen binding with equivalent sequences from human Fv variable regions. General methods for generating humanized antibodies are provided by Morrison (1985) Science 229:1202-1207, by Oi et al. (1986) BioTechniques 4:214, and by Queen et al. U.S. Pat. Nos. 5,585,089, 5,693,761 and 5,693,762, the contents of all of which are hereby incorporated by reference. Those methods include isolating, manipulating, and expressing the nucleic acid sequences that encode all or part of immunoglobulin Fv variable regions from at least one of a heavy or light chain. Sources of such nucleic acid are well known to those skilled in the art and, for example, may be obtained from a hybridoma producing an antibody against a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 polypeptide or fragment thereof. The recombinant DNA encoding the humanized antibody, or fragment thereof, can then be cloned into an appropriate expression vector.

Humanized or CDR-grafted antibodies can be produced by CDR-grafting or CDR substitution, wherein one, two, or all CDR's of an immunoglobulin chain can be replaced. See e.g., U.S. Pat. No. 5,225,539; Jones et al. (1986) Nature 321:552-525; Verhbeyan et al. (1988) Science 239:1534; Beidler et al. (1988) J. Immunol. 141:4053-4060; Winter U.S. Pat. No. 5,225,539, the contents of all of which are hereby expressly incorporated by reference. Winter describes a CDR-grafting method which may be used to prepare the humanized antibodies of the present invention (UK Patent Application GB 2188638A, filed on Mar. 26, 1987; Winter U.S. Pat. No. 5,225,539), the contents of which is expressly incorporated by reference.

Also within the scope of the invention are humanized antibodies in which specific amino acids have been substituted, deleted or added. Preferred humanized antibodies have amino acid substitutions in the framework region, such as to improve binding to the antigen. For example, a humanized antibody will have framework residues identical to the donor framework residue or to another amino acid other than the recipient framework residue. To generate such antibodies, a selected, small number of acceptor framework residues of the humanized immunoglobulin chain can be replaced by the corresponding donor amino acids. Preferred locations of the substitutions include amino acid residues adjacent to the CDR, or which are capable of interacting with a CDR (see e.g., U.S. Pat. No. 5,585,089). Criteria for selecting amino acids from the donor are described in U.S. Pat. No. 5,585,089, e.g., columns 12-16 of U.S. Pat. No. 5,585,089, the e.g., columns 12-16 of U.S. Pat. No. 5,585,089, the contents of which are hereby incorporated by reference. Other techniques for humanizing antibodies are described in Padlan et al. EP 519596 A1, published on Dec. 23, 1992.

Completely human antibodies are particularly desirable for therapeutic treatment of human patients. Such antibodies can be produced using transgenic mice that are incapable of expressing endogenous immunoglobulin heavy and light chains genes, but which can express human heavy and light chain genes. See, for example, Lonberg and Huszar (1995) Int. Rev. Immunol. 13:65-93); and U.S. Pat. Nos. 5,625,126; 5,633,425; 5,569,825; 5,661,016; and 5,545,806. In addition, companies such as Abgenix, Inc. (Fremont, Calif.) and Medarex, Inc. (Princeton, N.J.), can be engaged to provide human antibodies directed against a selected antigen using technology similar to that described above.

Completely human antibodies that recognize a selected epitope can be generated using a technique referred to as “guided selection.” In this approach a selected non-human monoclonal antibody, e.g., a murine antibody, is used to guide the selection of a completely human antibody recognizing the same epitope. This technology is described by Jespers et al. (1994) Bio/Technology 12:899-903).

The anti-26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 antibody can be a single chain antibody. A single-chain antibody (scFV) can be engineered as described in, for example, Colcher et al. (1999) Ann. N Y Acad. Sci. 880:263-80; and Reiter (1996) Clin. Cancer Res. 2:245-52. The single chain antibody can be dimerized or multimerized to generate multivalent antibodies having specificities for different epitopes of the same target 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein.

In a preferred embodiment, the antibody has reduced or no ability to bind an Fc receptor. For example, it is an isotype or subtype, fragment or other mutant, which does not support binding to an Fc receptor, e.g., it has a mutagenized or deleted Fc receptor binding region.

An antibody (or fragment thereof) may be conjugated to a therapeutic moiety such as a cytotoxin, a therapeutic agent or a radioactive ion. A cytotoxin or cytotoxic agent includes any agent that is detrimental to cells. Examples include taxol, cytochalasin B, gramicidin D, ethidium bromide, emetine, mitomycin, etoposide, tenoposide, vincristine, vinblastine, colchicin, doxorubicin, daunorubicin, dihydroxy anthracin dione, mitoxantrone, mithramycin, actinomycin D, 1-dehydrotestosterone, glucocorticoids, procaine, tetracaine, lidocaine, propranolol, puromycin, maytansinoids, e.g., maytansinol (see U.S. Pat. No. 5,208,020), CC-1065 (see U.S. Pat. Nos. 5,475,092, 5,585,499, 5,846,545) and analogs or homologs thereof. Therapeutic agents include, but are not limited to, antimetabolites (e.g., methotrexate, 6-mercaptopurine, 6-thioguanine, cytarabine, 5-fluorouracil decarbazine), alkylating agents (e.g., mechlorethamine, thioepa chlorambucil, CC-1065, melphalan, carmustine (BSNU) and lomustine (CCNU), cyclothosphamide, busulfan, dibromomannitol, streptozotocin, mitomycin C, and cis-dichlorodiamine platinum (II) (DDP) cisplatin), anthracyclines (e.g., daunorubicin (formerly daunomycin) and doxorubicin), antibiotics (e.g., dactinomycin (formerly actinomycin), bleomycin, mithramycin, and anthramycin (AMC)), and anti-mitotic agents (e.g., vincristine, vinblastine, taxol and maytansinoids).

Radioactive ions include, but are not limited to iodine, yttrium and praseodymium.

The conjugates of the invention can be used for modifying a given biological response, the therapeutic moiety is not to be construed as limited to classical chemical therapeutic agents. For example, the therapeutic moiety may be a protein or polypeptide possessing a desired biological activity. Such proteins may include, for example, a toxin such as abrin, ricin A, pseudomonas exotoxin, or diphtheria toxin; a protein such as tumor necrosis factor, α-interferon, β-interferon, nerve growth factor, platelet derived growth factor, tissue plasminogen activator; or, biological response modifiers such as, for example, lymphokines, interleukin-1 (“IL-1”), interleukin-2 (“IL-2”), interleukin-6 (“IL-6”), granulocyte macrophase colony stimulating factor (“GM-CSF”), granulocyte colony stimulating factor (“G-CSF”), or other growth factors.

Alternatively, an antibody can be conjugated to a second antibody to form an antibody heteroconjugate as described by Segal in U.S. Pat. No. 4,676,980.

An anti-26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 antibody (e.g., monoclonal antibody) can be used to isolate 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 by standard techniques, such as affinity chromatography or immunoprecipitation. Moreover, an anti-26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 antibody can be used to detect 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein (e.g., in a cellular lysate or cell supernatant) in order to evaluate the abundance and pattern of expression of the protein. Anti-26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 antibodies can be used diagnostically to monitor protein levels in tissue as part of a clinical testing procedure, e.g., to determine the efficacy of a given treatment regimen. Detection can be facilitated by coupling (i.e., physically linking) the antibody to a detectable substance (i.e., antibody labelling). Examples of detectable substances include various enzymes, prosthetic groups, fluorescent materials, luminescent materials, bioluminescent materials, and radioactive materials. Examples of suitable enzymes include horseradish peroxidase, alkaline phosphatase, β-galactosidase, or acetylcholinesterase; examples of suitable prosthetic group complexes include streptavidinibiotin and avidin/biotin; examples of suitable fluorescent materials include umbelliferone, fluorescein, fluorescein isothiocyanate, rhodamine, dichlorotriazinylamine fluorescein, dansyl chloride or phycoerythrin; an example of a luminescent material includes luminol; examples of bioluminescent materials include luciferase, luciferin, and aequorin, and examples of suitable radioactive material include ¹²⁵I, ¹³¹I, ³⁵S or ³H.

In preferred embodiments, an antibody can be made by immunizing with a purified 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 antigen, or a fragment thereof, e.g., a fragment described herein, a membrane associated antigen, tissues, e.g., crude tissue preparations, whole cells, preferably living cells, lysed cells, or cell fractions, e.g., membrane fractions.

Antibodies which bind only a native 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein, only denatured or otherwise non-native 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein, or which bind both, are within the invention. Antibodies with linear or conformational epitopes are within the invention. Conformational epitopes sometimes can be identified by identifying antibodies which bind to native but not denatured 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein.

Recombinant Expression Vectors, Host Cells and Genetically Engineered Cells

In another aspect, the invention includes, vectors, preferably expression vectors, containing a nucleic acid encoding a polypeptide described herein. As used herein, the term “vector” refers to a nucleic acid molecule capable of transporting another nucleic acid to which it has been linked and can include a plasmid, cosmid or viral vector. The vector can be capable of autonomous replication or it can integrate into a host DNA. Viral vectors include, e.g., replication defective retroviruses, adenoviruses and adeno-associated viruses.

A vector can include a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 nucleic acid in a form suitable for expression of the nucleic acid in a host cell.

Preferably the recombinant expression vector includes one or more regulatory sequences operatively linked to the nucleic acid sequence to be expressed. The term “regulatory sequence” includes promoters, enhancers and other expression control elements (e.g., polyadenylation signals). Regulatory sequences include those which direct constitutive expression of a nucleotide sequence, as well as tissue-specific regulatory and/or inducible sequences. The design of the expression vector can depend on such factors as the choice of the host cell to be transformed, the level of expression of protein desired, and the like. The expression vectors of the invention can be introduced into host cells to thereby produce proteins or polypeptides, including fusion proteins or polypeptides, encoded by nucleic acids as described herein (e.g., 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 proteins, mutant forms of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 proteins, fusion proteins, and the like).

The recombinant expression vectors of the invention can be designed for expression of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 proteins in prokaryotic or eukaryotic cells. For example, polypeptides of the invention can be expressed in E. coli, insect cells (e.g., using baculovirus expression vectors), yeast cells or mammalian cells. Suitable host cells are discussed further in Goeddel, (1990) Gene Expression Technology: Methods in Enzymology 185, Academic Press, San Diego, Calif. Alternatively, the recombinant expression vector can be transcribed and translated in vitro, for example using T7 promoter regulatory sequences and T7 polymerase.

Expression of proteins in prokaryotes is most often carried out in E. coli with vectors containing constitutive or inducible promoters directing the expression of either fusion or non-fusion proteins. Fusion vectors add a number of amino acids to a protein encoded therein, usually to the amino terminus of the recombinant protein. Such fusion vectors typically serve three purposes: 1) to increase expression of recombinant protein; 2) to increase the solubility of the recombinant protein; and 3) to aid in the purification of the recombinant protein by acting as a ligand in affinity purification. Often, a proteolytic cleavage site is introduced at the junction of the fusion moiety and the recombinant protein to enable separation of the recombinant protein from the fusion moiety subsequent to purification of the fusion protein. Such enzymes, and their cognate recognition sequences, include Factor Xa, thrombin and enterokinase. Typical fusion expression vectors include pGEX (Pharmacia Biotech Inc; Smith and Johnson (1988) Gene 67:31-40), pMAL (New England Biolabs, Beverly, Mass.) and pRIT5 (Pharmacia, Piscataway, N.J.) which fuse glutathione S-transferase (GST), maltose E binding protein, or protein A, respectively, to the target recombinant protein.

Purified fusion proteins can be used in 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 activity assays, (e.g., direct assays or competitive assays described in detail below), or to generate antibodies specific or selective for 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 proteins. In a preferred embodiment, a fusion protein expressed in a retroviral expression vector of the present invention can be used to infect bone marrow cells which are subsequently transplanted into irradiated recipients. The pathology of the subject recipient is then examined after sufficient time has passed (e.g., six weeks).

To maximize recombinant protein expression in E. coli is to express the protein in a host bacteria with an impaired capacity to proteolytically cleave the recombinant protein (Gottesman (1990) Gene Expression Technology: Methods in Enzymology 185, Academic Press, San Diego, Calif. 119-128). Another strategy is to alter the nucleic acid sequence of the nucleic acid to be inserted into an expression vector so that the individual codons for each amino acid are those preferentially utilized in E. coli (Wada et al., (1992) Nucleic Acids Res. 20:2111-2118). Such alteration of nucleic acid sequences of the invention can be carried out by standard DNA synthesis techniques.

The 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 expression vector can be a yeast expression vector, a vector for expression in insect cells, e.g., a baculovirus expression vector or a vector suitable for expression in mammalian cells.

When used in mammalian cells, the expression vector's control functions are often provided by viral regulatory elements. For example, commonly used promoters are derived from polyoma, Adenovirus 2, cytomegalovirus and Simian Virus 40.

In another embodiment, the recombinant mammalian expression vector is capable of directing expression of the nucleic acid preferentially in a particular cell type (e.g., tissue-specific regulatory elements are used to express the nucleic acid). Non-limiting examples of suitable tissue-specific promoters include the albumin promoter (liver-specific; Pinkert et al. (1987) Genes Dev. 1:268-277), lymphoid-specific promoters (Calame and Eaton (1988) Adv. Immunol. 43:235-275), in particular promoters of T cell receptors (Winoto and Baltimore (1989) EMBO J. 8:729-733) and immunoglobulins (Banerji et al. (1983) Cell 33:729-740; Queen and Baltimore (1983) Cell 33:741-748), neuron-specific promoters (e.g., the neurofilament promoter; Byrne and Ruddle (1989) Proc. Natl. Acad. Sci. USA 86:5473-5477), pancreas-specific promoters (Edlund et al. (1985) Science 230:912-916), and mammary gland-specific promoters (e.g., milk whey promoter; U.S. Pat. No. 4,873,316 and European Application Publication No. 264,166). Developmentally-regulated promoters are also encompassed, for example, the murine hox promoters (Kessel and Gruss (1990) Science 249:374-379) and the α-fetoprotein promoter (Campes and Tilghman (1989) Genes Dev. 3:537-546).

The invention further provides a recombinant expression vector comprising a DNA molecule of the invention cloned into the expression vector in an antisense orientation. Regulatory sequences (e.g., viral promoters and/or enhancers) operatively linked to a nucleic acid cloned in the antisense orientation can be chosen which direct the constitutive, tissue specific or cell type specific expression of antisense RNA in a variety of cell types. The antisense expression vector can be in the form of a recombinant plasmid, phagemid or attenuated virus. For a discussion of the regulation of gene expression using antisense genes see Weintraub et al., (1986) Reviews—Trends in Genetics 1:1.

Another aspect the invention provides a host cell which includes a nucleic acid molecule described herein, e.g., a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 nucleic acid molecule within a recombinant expression vector or a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 nucleic acid molecule containing sequences which allow it to homologously recombine into a specific site of the host cell's genome. The terms “host cell” and “recombinant host cell” are used interchangeably herein. Such terms refer not only to the particular subject cell but to the progeny or potential progeny of such a cell. Because certain modifications can occur in succeeding generations due to either mutation or environmental influences, such progeny may not, in fact, be identical to the parent cell, but are still included within the scope of the term as used herein.

A host cell can be any prokaryotic or eukaryotic cell. For example, a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein can be expressed in bacterial cells such as E. coli, insect cells, yeast or mammalian cells (such as Chinese hamster ovary (CHO) cells or CV-1 origin, SV-40 (COS) cells). Other suitable host cells are known to those skilled in the art.

Vector DNA can be introduced into host cells via conventional transformation or transfection techniques. As used herein, the terms “transformation” and “transfection” are intended to refer to a variety of art-recognized techniques for introducing foreign nucleic acid (e.g., DNA) into a host cell, including calcium phosphate or calcium chloride co-precipitation, DEAE-dextran-mediated transfection, lipofection, or electroporation.

A host cell of the invention can be used to produce (i.e., express) a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein. Accordingly, the invention further provides methods for producing a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein using the host cells of the invention. In one embodiment, the method includes culturing the host cell of the invention (into which a recombinant expression vector encoding a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein has been introduced) in a suitable medium such that a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein is produced. In another embodiment, the method further includes isolating a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein from the medium or the host cell.

In another aspect, the invention features, a cell or purified preparation of cells which include a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 transgene, or which otherwise misexpress 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843. The cell preparation can consist of human or non-human cells, e.g., rodent cells, e.g., mouse or rat cells, rabbit cells, or pig cells. In preferred embodiments, the cell or cells include a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 transgene, e.g., a heterologous form of a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843, e.g., a gene derived from humans (in the case of a non-human cell). The 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 transgene can be misexpressed, e.g., overexpressed or underexpressed. In other preferred embodiments, the cell or cells include a gene which misexpresses an endogenous 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843, e.g., a gene the expression of which is disrupted, e.g., a knockout. Such cells can serve as a model for studying disorders which are related to mutated or misexpressed 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 alleles or for use in drug screening.

In another aspect, the invention features, a human cell, e.g., a hematopoietic stem cell, transformed with nucleic acid which encodes a subject 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 polypeptide.

Also provided are cells, preferably human cells, e.g., human hematopoietic or fibroblast cells, in which an endogenous 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 is under the control of a regulatory sequence that does not normally control the expression of the endogenous 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 gene. The expression characteristics of an endogenous gene within a cell, e.g., a cell line or microorganism, can be modified by inserting a heterologous DNA regulatory element into the genome of the cell such that the inserted regulatory element is operably linked to the endogenous 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 gene. For example, an endogenous 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 gene which is “transcriptionally silent,” e.g., not normally expressed, or expressed only at very low levels, can be activated by inserting a regulatory element which is capable of promoting the expression of a normally expressed gene product in that cell. Techniques such as targeted homologous recombinations, can be used to insert the heterologous DNA as described in, e.g., Chappel, U.S. Pat. No. 5,272,071; WO 91/06667, published in May 16, 1991.

Transgenic Animals

The invention provides non-human transgenic animals. Such animals are useful for studying the function and/or activity of a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein and for identifying and/or evaluating modulators of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 activity. As used herein, a “transgenic animal” is a non-human animal, preferably a mammal, more preferably a rodent such as a rat or mouse, in which one or more of the cells of the animal includes a transgene. Other examples of transgenic animals include non-human primates, sheep, dogs, cows, goats, chickens, amphibians, and the like. A transgene is exogenous DNA or a rearrangement, e.g., a deletion of endogenous chromosomal DNA, which preferably is integrated into or occurs in the genome of the cells of a transgenic animal. A transgene can direct the expression of an encoded gene product in one or more cell types or tissues of the transgenic animal, other transgenes, e.g., a knockout, reduce expression. Thus, a transgenic animal can be one in which an endogenous 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 gene has been altered by, e.g., by homologous recombination between the endogenous gene and an exogenous DNA molecule introduced into a cell of the animal, e.g., an embryonic cell of the animal, prior to development of the animal.

Intronic sequences and polyadenylation signals can also be included in the transgene to increase the efficiency of expression of the transgene. A tissue-specific regulatory sequence(s) can be operably linked to a transgene of the invention to direct expression of a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein to particular cells. A transgenic founder animal can be identified based upon the presence of a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 transgene in its genome and/or expression of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 mRNA in tissues or cells of the animals. A transgenic founder animal can then be used to breed additional animals carrying the transgene. Moreover, transgenic animals carrying a transgene encoding a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein can further be bred to other transgenic animals carrying other transgenes.

26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 proteins or polypeptides can be expressed in transgenic animals or plants, e.g., a nucleic acid encoding the protein or polypeptide can be introduced into the genome of an animal. In preferred embodiments the nucleic acid is placed under the control of a tissue specific promoter, e.g., a milk or egg specific promoter, and recovered from the milk or eggs produced by the animal. Suitable animals are mice, pigs, cows, goats, and sheep.

The invention also includes a population of cells from a transgenic animal, as discussed, e.g., below.

Uses

The nucleic acid molecules, proteins, protein homologs, and antibodies described herein can be used in one or more of the following methods: a) screening assays; b) predictive medicine (e.g., diagnostic assays, prognostic assays, monitoring clinical trials, and pharmacogenetics); and c) methods of treatment (e.g., therapeutic and prophylactic).

The isolated nucleic acid molecules of the invention can be used, for example, to express a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein (e.g., via a recombinant expression vector in a host cell in gene therapy applications), to detect a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 mRNA (e.g., in a biological sample) or a genetic alteration in a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 gene, and to modulate 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 activity, as described further below. The 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 proteins can be used to treat disorders characterized by insufficient or excessive production of a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 substrate or production of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 inhibitors. In addition, the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 proteins can be used to screen for naturally occurring 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 substrates, to screen for drugs or compounds which modulate 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 activity, as well as to treat disorders characterized by insufficient or excessive production of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein or production of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein forms which have decreased, aberrant or unwanted activity compared to 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 wild type protein (e.g., aberrant or deficient arginine methyltransferase activity, glycosyltransferase activity, gamma-glutamyltraspeptidase activity, phosphoribosylglycinamide transferase activity, acyltransferase activity, acyl-CoA dehydrogenase activity, fatty acid amide hydrolase activity, aminotransferase activity, zinc carboxypeptidase activity, protein kinase activity, DEAD helicase activity, short-chain dehydrogenase/reductase activity or phosphatase activity, or other activity). Moreover, the anti-26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 antibodies of the invention can be used to detect and isolate 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 proteins, regulate the bioavailability of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 proteins, and modulate 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 activity.

A method of evaluating a compound for the ability to interact with, e.g., bind, a subject 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 polypeptide is provided. The method includes: contacting the compound with the subject 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 polypeptide; and evaluating ability of the compound to interact with, e.g., to bind or form a complex with the subject 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 polypeptide. This method can be performed in vitro, e.g., in a cell free system, or in vivo, e.g., in a two-hybrid interaction trap assay. This method can be used to identify naturally occurring molecules which interact with subject 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 polypeptide. It can also be used to find natural or synthetic inhibitors of subject 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 polypeptide. Screening methods are discussed in more detail below.

Screening Assays

The invention provides methods (also referred to herein as “screening assays”) for identifying modulators, i.e., candidate or test compounds or agents (e.g., proteins, peptides, peptidomimetics, peptoids, small molecules or other drugs) which bind to 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 proteins, have a stimulatory or inhibitory effect on, for example, 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 expression or 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 activity, or have a stimulatory or inhibitory effect on, for example, the expression or activity of a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 substrate. Compounds thus identified can be used to modulate the activity of target gene products (e.g., 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 genes) in a therapeutic protocol, to elaborate the biological function of the target gene product, or to identify compounds that disrupt normal target gene interactions.

In one embodiment, the invention provides assays for screening candidate or test compounds which are substrates of a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein or polypeptide or a biologically active portion thereof. In another embodiment, the invention provides assays for screening candidate or test compounds which bind to or modulate the activity of a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein or polypeptide or a biologically active portion thereof.

The test compounds of the present invention can be obtained using any of the numerous approaches in combinatorial library methods known in the art, including: biological libraries; peptoid libraries (libraries of molecules having the functionalities of peptides, but with a novel, non-peptide backbone which are resistant to enzymatic degradation but which nevertheless remain bioactive; see, e.g., Zuckermann et al. (1994) J. Med. Chem. 37:2678-85); spatially addressable parallel solid phase or solution phase libraries; synthetic library methods requiring deconvolution; the ‘one-bead one-compound’ library method; and synthetic library methods using affinity chromatography selection. The biological library and peptoid library approaches are limited to peptide libraries, while the other four approaches are applicable to peptide, non-peptide oligomer or small molecule libraries of compounds (Lam (1997) Anticancer Drug Des. 12:145).

Examples of methods for the synthesis of molecular libraries can be found in the art, for example in: DeWitt et al. (1993) Proc. Natl. Acad. Sci. U.S.A. 90:6909-13; Erb et al. (1994) Proc. Natl. Acad. Sci. USA 91:11422-426; Zuckermann et al. (1994). J. Med. Chem. 37:2678-85; Cho et al. (1993) Science 261:1303; Carrell et al. (1994) Angew. Chem. Int. Ed. Engl. 33:2059; Carell et al. (1994) Angew. Chem. Int. Ed. Engl. 33:2061; and in Gallop et al. (1994) J. Med. Chem. 37:1233-51.

Libraries of compounds can be presented in solution (e.g., Houghten (1992) Biotechniques 13:412-421), or on beads (Lam (1991) Nature 354:82-84), chips (Fodor (1993) Nature 364:555-556), bacteria (Ladner, U.S. Pat. No. 5,223,409), spores (Ladner U.S. Pat. No. '409), plasmids (Cull et al. (1992) Proc Natl Acad Sci USA 89:1865-1869) or on phage (Scott and Smith (1990) Science 249:386-390; Devlin (1990) Science 249:404-406; Cwirla et al. (1990) Proc. Natl. Acad. Sci. 87:6378-6382; Felici (1991) J. Mol. Biol. 222:301-310; Ladner supra.).

In one embodiment, an assay is a cell-based assay in which a cell which expresses a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein or biologically active portion thereof is contacted with a test compound, and the ability of the test compound to modulate 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 activity is determined. Determining the ability of the test compound to modulate 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 activity can be accomplished by monitoring, for example, arginine methyltransferase activity, glycosyltransferase activity, gamma-glutamyltraspeptidase activity, phosphoribosylglycinamide transferase activity, acyltransferase activity, acyl-CoA dehydrogenase activity, fatty acid amide hydrolase activity, aminotransferase activity, zinc carboxypeptidase activity, protein kinase activity, DEAD helicase activity, short-chain dehydrogenase/reductase activity or phosphatase activity, or other activity. The cell, for example, can be of mammalian origin, e.g., human.

The ability of the test compound to modulate 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 binding to a compound, e.g., a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 substrate, or to bind to 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 can also be evaluated. This can be accomplished, for example, by coupling the compound, e.g., the substrate, with a radioisotope or enzymatic label such that binding of the compound, e.g., the substrate, to 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 can be determined by detecting the labeled compound, e.g., substrate, in a complex. Alternatively, 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 could be coupled with a radioisotope or enzymatic label to monitor the ability of a test compound to modulate 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 binding to a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 substrate in a complex. For example, compounds (e.g., 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 substrates) can be labeled with ¹²⁵I, ¹⁴C, ³⁵S or ³H., either directly or indirectly, and the radioisotope detected by direct counting of radioemmission or by scintillation counting. Alternatively, compounds can be enzymatically labeled with, for example, horseradish peroxidase, alkaline phosphatase, or luciferase, and the enzymatic label detected by determination of conversion of an appropriate substrate to product.

The ability of a compound (e.g., a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 substrate) to interact with 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 with or without the labeling of any of the interactants can be evaluated. For example, a microphysiometer can be used to detect the interaction of a compound with 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 without the labeling of either the compound or the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843. McConnell et al. (1992) Science 257:1906-1912. As used herein, a “microphysiometer” (e.g., Cytosensor) is an analytical instrument that measures the rate at which a cell acidifies its environment using a light-addressable potentiometric sensor (LAPS). Changes in this acidification rate can be used as an indicator of the interaction between a compound and 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843.

In yet another embodiment, a cell-free assay is provided in which a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein or biologically active portion thereof is contacted with a test compound and the ability of the test compound to bind to the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein or biologically active portion thereof is evaluated. Preferred biologically active portions of the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 proteins to be used in assays of the present invention include fragments which participate in interactions with non-26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 molecules, e.g., fragments with high surface probability scores.

Soluble and/or membrane-bound forms of isolated proteins (e.g., 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 proteins or biologically active portions thereof) can be used in the cell-free assays of the invention. When membrane-bound forms of the protein are used, it may be desirable to utilize a solubilizing agent. Examples of such solubilizing agents include non-ionic detergents such as n-octylglucoside, n-dodecylglucoside, n-dodecylmaltoside, octanoyl-N-methylglucamide, decanoyl-N-methylglucamide, Triton® X-100, Triton® X-114, Thesit®, Isotridecypoly(ethylene glycol ether) _n, 3-[(3-cholamidopropyl)dimethylamminio]-1-propane sulfonate (CHAPS), 3-[(3-cholamidopropyl)dimethylamminio]-2-hydroxy-1-propane sulfonate (CHAPSO), or N-dodecyl═N,N-dimethyl-3-ammonio-1-propane sulfonate.

Cell-free assays involve preparing a reaction mixture of the target gene protein and the test compound under conditions and for a time sufficient to allow the two components to interact and bind, thus forming a complex that can be removed and/or detected.

The interaction between two molecules can also be detected, e.g., using fluorescence energy transfer (FET) (see, for example, Lakowicz et al., U.S. Pat. No. 5,631,169; Stavrianopoulos, et al., U.S. Pat. No. 4,868,103). A fluorophore label on the first, ‘donor’ molecule is selected such that its emitted fluorescent energy will be absorbed by a fluorescent label on a second, ‘acceptor’ molecule, which in turn is able to fluoresce due to the absorbed energy. Alternately, the ‘donor’ protein molecule can simply utilize the natural fluorescent energy of tryptophan residues. Labels are chosen that emit different wavelengths of light, such that the ‘acceptor’ molecule label can be differentiated from that of the ‘donor’. Since the efficiency of energy transfer between the labels is related to the distance separating the molecules, the spatial relationship between the molecules can be assessed. In a situation in which binding occurs between the molecules, the fluorescent emission of the ‘acceptor’ molecule label in the assay should be maximal. An FET binding event can be conveniently measured through standard fluorometric detection means well known in the art (e.g., using a fluorimeter).

In another embodiment, determining the ability of the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein to bind to a target molecule can be accomplished using real-time Biomolecular Interaction Analysis (BIA) (see, e.g., Sjolander and Urbaniczky (1991) Anal. Chem. 63:2338-2345 and Szabo et al. (1995) Curr. Opin. Struct. Biol. 5:699-705). “Surface plasmon resonance” or “BIA” detects biospecific interactions in real time, without labeling any of the interactants (e.g., BIAcore). Changes in the mass at the binding surface (indicative of a binding event) result in alterations of the refractive index of light near the surface (the optical phenomenon of surface plasmon resonance (SPR)), resulting in a detectable signal which can be used as an indication of real-time reactions between biological molecules.

In one embodiment, the target gene product or the test substance is anchored onto a solid phase. The target gene product/test compound complexes anchored on the solid phase can be detected at the end of the reaction. Preferably, the target gene product can be anchored onto a solid surface, and the test compound, (which is not anchored), can be labeled, either directly or indirectly, with detectable labels discussed herein.

It may be desirable to immobilize either 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843, an anti-26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 antibody or its target molecule to facilitate separation of complexed from uncomplexed forms of one or both of the proteins, as well as to accommodate automation of the assay. Binding of a test compound to a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein, or interaction of a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein with a target molecule in the presence and absence of a candidate compound, can be accomplished in any vessel suitable for containing the reactants. Examples of such vessels include microtiter plates, test tubes, and micro-centrifuge tubes. In one embodiment, a fusion protein can be provided which adds a domain that allows one or both of the proteins to be bound to a matrix. For example, glutathione-S-transferase/26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 fusion proteins or glutathione-S-transferase/target fusion proteins can be adsorbed onto glutathione sepharose beads (Sigma Chemical, St. Louis, Mo.) or glutathione derivatized microtiter plates, which are then combined with the test compound or the test compound and either the non-adsorbed target protein or 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein, and the mixture incubated under conditions conducive to complex formation (e.g., at physiological conditions for salt and pH).

Following incubation, the beads or microtiter plate wells are washed to remove any unbound components, the matrix immobilized in the case of beads, complex determined either directly or indirectly, for example, as described above. Alternatively, the complexes can be dissociated from the matrix, and the level of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 binding or activity determined using standard techniques.

Other techniques for immobilizing either a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein or a target molecule on matrices include using conjugation of biotin and streptavidin. Biotinylated 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein or target molecules can be prepared from biotin-NHS (N-hydroxy-succinimide) using techniques known in the art (e.g., biotinylation kit, Pierce Chemicals, Rockford, Ill.), and immobilized in the wells of streptavidin-coated 96 well. plates (Pierce Chemical).

In order to conduct the assay, the non-immobilized component is added to the coated surface containing the anchored component. After the reaction is complete, unreacted components are removed (e.g., by washing) under conditions such that any complexes formed will remain immobilized on the solid surface. The detection of complexes anchored on the solid surface can be accomplished in a number of ways. Where the previously non-immobilized component is pre-labeled, the detection of label immobilized on the surface indicates that complexes were formed. Where the previously non-immobilized component is not pre-labeled, an indirect label can be used to detect complexes anchored on the surface; e.g., using a labeled antibody specific or selective for the immobilized component (the antibody, in turn, can be directly labeled or indirectly labeled with, e.g., a labeled anti-Ig antibody).

In one embodiment, this assay is performed utilizing antibodies reactive with 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein or target molecules but which do not interfere with binding of the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein to its target molecule. Such antibodies can be derivatized to the wells of the plate, and unbound target or 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein trapped in the wells by antibody conjugation. Methods for detecting such complexes, in addition to those described above for the GST-immobilized complexes, include immunodetection of complexes using antibodies reactive with the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein or target molecule, as well as enzyme-linked assays which rely on detecting an enzymatic activity associated with the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein or target molecule.

Alternatively, cell free assays can be conducted in a liquid phase. In such an assay, the reaction products are separated from unreacted components, by any of a number of standard techniques, including but not limited to: differential centrifugation (see, for example, Rivas and Minton (1993) Trends Biochem Sci 18:284-7); chromatography (gel filtration chromatography, ion-exchange chromatography); electrophoresis (see, e.g., Ausubel et al., eds. (1999) Current Protocols in Molecular Biology, J. Wiley, New York.); and immunoprecipitation (see, for example, Ausubel et al., eds. (1999) Current Protocols in Molecular Biology, J. Wiley, New York). Such resins and chromatographic techniques are known to one skilled in the art (see, e.g., Heegaard (1998) J Mol Recognit 11: 141-8; Hage and Tweed (1997) J Chromatogr B Biomed Sci Appl. 699:499-525). Further, fluorescence energy transfer can also be conveniently utilized, as described herein, to detect binding without further purification of the complex from solution.

In a preferred embodiment, the assay includes contacting the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein or biologically active portion thereof with a known compound which binds 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 to form an assay mixture, contacting the assay mixture with a test compound, and determining the ability of the test compound to interact with a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein, wherein determining the ability of the test compound to interact with a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein includes determining the ability of the test compound to preferentially bind to 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 or biologically active portion thereof, or to modulate the activity of a target molecule, as compared to the known compound.

The target gene products of the invention can, in vivo, interact with one or more cellular or extracellular macromolecules, such as proteins. For the purposes of this discussion, such cellular and extracellular macromolecules are referred to herein as “binding partners.” Compounds that disrupt such interactions can be useful in regulating the activity of the target gene product. Such compounds can include, but are not limited to molecules such as antibodies, peptides, and small molecules. The preferred target genes/products for use in this embodiment are the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 genes herein identified. In an alternative embodiment, the invention provides methods for determining the ability of the test compound to modulate the activity of a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein through modulation of the activity of a downstream effector of a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 target molecule. For example, the activity of the effector molecule on an appropriate target can be determined, or the binding of the effector to an appropriate target can be determined, as previously described.

To identify compounds that interfere with the interaction between the target gene product and its cellular or extracellular binding partner(s), a reaction mixture containing the target gene product and the binding partner is prepared, under conditions and for a time sufficient, to allow the two products to form complex. In order to test an inhibitory agent, the reaction mixture is provided in the presence and absence of the test compound. The test compound can be initially included in the reaction mixture, or can be added at a time subsequent to the addition of the target gene and its cellular or extracellular binding partner. Control reaction mixtures are incubated without the test compound or with a placebo. The formation of any complexes between the target gene product and the cellular or extracellular binding partner is then detected. The formation of a complex in the control reaction, but not in the reaction mixture containing the test compound, indicates that the compound interferes with the interaction of the target gene product and the interactive binding partner.

Additionally, complex formation within reaction mixtures containing the test compound and normal target gene product can also be compared to complex formation within reaction mixtures containing the test compound and mutant target gene product. This comparison can be important in those cases wherein it is desirable to identify compounds that disrupt interactions of mutant but not normal target gene products.

These assays can be conducted in a heterogeneous or homogeneous format. Heterogeneous assays involve anchoring either the target gene product or the binding partner onto a solid phase, and detecting complexes anchored on the solid phase at the end of the reaction. In homogeneous assays, the entire reaction is carried out in a liquid phase. In either approach, the order of addition of reactants can be varied to obtain different information about the compounds being tested. For example, test compounds that interfere with the interaction between the target gene products and the binding partners, e.g., by competition, can be identified by conducting the reaction in the presence of the test substance. Alternatively, test compounds that disrupt preformed complexes, e.g., compounds with higher binding constants that displace one of the components from the complex, can be tested by adding the test compound to the reaction mixture after complexes have been formed. The various formats are briefly described below.

In a heterogeneous assay system, either the target gene product or the interactive cellular or extracellular binding partner, is anchored onto a solid surface (e.g., a microtiter plate), while the non-anchored species is labeled, either directly or indirectly. The anchored species can be immobilized by non-covalent or covalent attachments. Alternatively, an immobilized antibody specific or selective for the species to be anchored can be used to anchor the species to the solid surface.

In order to conduct the assay, the partner of the immobilized species is exposed to the coated surface with or without the test compound. After the reaction is complete, unreacted components are removed (e.g., by washing) and any complexes formed will remain immobilized on the solid surface. Where the non-immobilized species is pre-labeled, the detection of label immobilized on the surface indicates that complexes were formed. Where the non-immobilized species is not pre-labeled, an indirect label can be used to detect complexes anchored on the surface; e.g., using a labeled antibody specific or selective for the initially non-immobilized species (the antibody, in turn, can be directly labeled or indirectly labeled with, e.g., a labeled anti-Ig antibody). Depending upon the order of addition of reaction components, test compounds that inhibit complex formation or that disrupt preformed complexes can be detected.

Alternatively, the reaction can be conducted in a liquid phase in the presence or absence of the test compound, the reaction products separated from unreacted components, and complexes detected; e.g., using an immobilized antibody specific or selective for one of the binding components to anchor any complexes formed in solution, and a labeled antibody specific or selective for the other partner to detect anchored complexes. Again, depending upon the order of addition of reactants to the liquid phase, test compounds that inhibit complex or that disrupt preformed complexes can be identified.

In an alternate embodiment of the invention, a homogeneous assay can be used. For example, a preformed complex of the target gene product and the interactive cellular or extracellular binding partner product is prepared in that either the target gene products or their binding partners are labeled, but the signal generated by the label is quenched due to complex formation (see, e.g., U.S. Pat. No. 4,109,496 that utilizes this approach for immunoassays). The addition of a test substance that competes with and displaces one of the species from the preformed complex will result in the generation of a signal above background. In this way, test substances that disrupt target gene product-binding partner interaction can be identified.

In yet another aspect, the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 proteins can be used as “bait proteins” in a two-hybrid assay or three-hybrid assay (see, e.g., U.S. Pat. No. 5,283,317; Zervos et al. (1993) Cell 72:223-232; Madura et al. (1993) J. Biol. Chem. 268:12046-12054; Bartel et al. (1993) Biotechniques 14:920-924; Iwabuchi et al. (1993) Oncogene 8:1693-1696; and Brent WO94/10300), to identify other proteins, which bind to or interact with 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 (“26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843-binding proteins” or “26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843-bp”) and are involved in 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 activity. Such 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843-bps can be activators or inhibitors of signals by the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 proteins or 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 targets as, for example, downstream elements of a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843-mediated signaling pathway.

The two-hybrid system is based on the modular nature of most transcription factors, which consist of separable DNA-binding and activation domains. Briefly, the assay utilizes two different DNA constructs. In one construct, the gene that codes for a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein is fused to a gene encoding the DNA binding domain of a known transcription factor (e.g., GAL-4). In the other construct, a DNA sequence, from a library of DNA sequences, that encodes an unidentified protein (“prey” or “sample”) is fused to a gene that codes for the activation domain of the known transcription factor. (Alternatively the: 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein can be the fused to the activator domain.) If the “bait” and the “prey” proteins are able to interact, in vivo, forming a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843-dependent complex, the DNA-binding and activation domains of the transcription factor are brought into close proximity. This proximity allows transcription of a reporter gene (e.g., lacZ) which is operably linked to a transcriptional regulatory site responsive to the transcription factor. Expression of the reporter gene can be detected and cell colonies containing the functional transcription factor can be isolated and used to obtain the cloned gene which encodes the protein which interacts with the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein.

In another embodiment, modulators of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 expression are identified. For example, a cell or cell free mixture is contacted with a candidate compound and the expression of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 mRNA or protein evaluated relative to the level of expression of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 mRNA or protein in the absence of the candidate compound. When expression of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 mRNA or protein is greater in the presence of the candidate compound than in its absence, the candidate compound is identified as a stimulator of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 mRNA or protein expression. Alternatively, when expression of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 mRNA or protein is less (statistically significantly less) in the presence of the candidate compound than in its absence, the candidate compound is identified as an inhibitor of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 mRNA or protein expression. The level of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 mRNA or protein expression can be determined by methods described herein for detecting 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 mRNA or protein.

In another aspect, the invention pertains to a combination of two or more of the assays described herein. For example, a modulating agent can be identified using a cell-based or a cell free assay, and the ability of the agent to modulate the activity of a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein can be confirmed in vivo, e.g., in an animal such as an animal model for aberrant or deficient arginine methyltransferase activity, glycosyltransferase activity, gamma-glutamyltraspeptidase activity, phosphoribosylglycinamide transferase activity, acyltransferase activity, acyl-CoA dehydrogenase activity, fatty acid amide hydrolase activity, aminotransferase activity, zinc carboxypeptidase activity, protein kinase activity, DEAD helicase activity, short-chain dehydrogenase/reductase activity or phosphatase activity.

This invention further pertains to novel agents identified by the above-described screening assays. Accordingly, it is within the scope of this invention to further use an agent identified as described herein (e.g., a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 modulating agent, an antisense 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 nucleic acid molecule, a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843-specific antibody, or a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843-binding partner) in an appropriate animal model to determine the efficacy, toxicity, side effects, or mechanism of action, of treatment with such an agent. Furthermore, novel agents identified by the above-described screening assays can be used for treatments as described herein.

Detection Assays

Portions or fragments of the nucleic acid sequences identified herein can be used as polynucleotide reagents. For example, these sequences can be used to: (i) map their respective genes on a chromosome e.g., to locate gene regions associated with genetic disease or to associate 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 with a disease; (ii) identify an individual from a minute biological sample (tissue typing); and (iii) aid in forensic identification of a biological sample. These applications are described in the subsections below.

Chromosome Mapping

The 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 nucleotide sequences or portions thereof can be used to map the location of the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 genes on a chromosome. This process is called chromosome mapping. Chromosome mapping is useful in correlating the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 sequences with genes associated with disease.

Briefly, 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 genes can be mapped to chromosomes by preparing PCR primers (preferably 15-25 bp in length) from the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 nucleotide sequences. These primers can then be used for PCR screening of somatic cell hybrids containing individual human chromosomes. Only those hybrids containing the human gene corresponding to the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 sequences will yield an amplified fragment.

A panel of somatic cell hybrids in which each cell line contains either a single human chromosome or a small number of human chromosomes, and a full set of mouse chromosomes, can allow easy mapping of individual genes to specific human chromosomes. (D'Eustachio et al. (1983) Science 220:919-924).

Other mapping strategies e.g., in situ hybridization (described in Fan et al. (1990) Proc. Natl. Acad. Sci. USA, 87:6223-27), pre-screening with labeled flow-sorted chromosomes, and pre-selection by hybridization to chromosome specific cDNA libraries can be used to map 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 to a chromosomal location.

Fluorescence in situ hybridization (FISH) of a DNA sequence to a metaphase chromosomal spread can further be used to provide a precise chromosomal location in one step. The FISH technique can be used with a DNA sequence as short as 500 or 600 bases. However, clones larger than 1,000 bases have a higher likelihood of binding to a unique chromosomal location with sufficient signal intensity for simple detection. Preferably 1,000 bases, and more preferably 2,000 bases will suffice to get good results at a reasonable amount of time. For a review of this technique, see Verma et al. (1988) Human Chromosomes: A Manual of Basic Techniques, Pergamon Press, New York).

Reagents for chromosome mapping can be used individually to mark a single chromosome or a single site on that chromosome, or panels of reagents can be used for marking multiple sites and/or multiple chromosomes. Reagents corresponding to noncoding regions of the genes actually are preferred for mapping purposes. Coding sequences are more likely to be conserved within gene families, thus increasing the chance of cross hybridizations during chromosomal mapping.

Once a sequence has been mapped to a precise chromosomal location, the physical position of the sequence on the chromosome can be correlated with genetic map data. (Such data are found, for example, in McKusick, Mendelian Inheritance in Man, available on-line through Johns Hopkins University Welch Medical Library). The relationship between a gene and a disease, mapped to the same chromosomal region, can then be identified through linkage analysis (co-inheritance of physically adjacent genes), described in, for example, Egeland et al. (1987) Nature, 325:783-787.

Moreover, differences in the DNA sequences between individuals affected and unaffected with a disease associated with the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 gene, can be determined. If a mutation is observed in some or all of the affected individuals but not in any unaffected individuals, then the mutation is likely to be the causative agent of the particular disease. Comparison of affected and unaffected individuals generally involves first looking for structural alterations in the chromosomes, such as deletions or translocations that are visible from chromosome spreads or detectable using PCR based on that DNA sequence. Ultimately, complete sequencing of genes from several individuals can be performed to confirm the presence of a mutation and to distinguish mutations from polymorphisms.

Tissue Typing

26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 sequences can be used to identify individuals from biological samples using, e.g., restriction fragment length polymorphism (RFLP). In this technique, an individual's genomic DNA is digested with one or more restriction enzymes, the fragments separated, e.g., in a Southern blot, and probed to yield bands for identification. The sequences of the present invention are useful as additional DNA markers for RFLP (described in U.S. Pat. No. 5,272,057).

Furthermore, the sequences of the present invention can also be used to determine the actual base-by-base DNA sequence of selected portions of an individual's genome. Thus, the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 nucleotide sequences described herein can be used to prepare two PCR primers from the 5′ and 3′ ends of the sequences. These primers can then be used to amplify an individual's DNA and subsequently sequence it. Panels of corresponding DNA sequences from individuals, prepared in this manner, can provide unique individual identifications, as each individual will have a unique set of such DNA sequences due to allelic differences.

Allelic variation occurs to some degree in the coding regions of these sequences, and to a greater degree in the noncoding regions. Each of the sequences described herein can, to some degree, be used as a standard against which DNA from an individual can be compared for identification purposes. Because greater numbers of polymorphisms occur in the noncoding regions, fewer sequences are necessary to differentiate individuals. The noncoding sequences of SEQ ID NO:1, 4, 7, 10, 13, 16, 53, 61, 67, 78, 88, 100, 113, 122 or 129 can provide positive individual identification with a panel of perhaps 10 to 1,000 primers which each yield a noncoding amplified sequence of 100 bases. If predicted coding sequences, such as those in SEQ ID NO:3, 6, 9, 12, 15, 18, 55, 63, 69, 80, 90, 102, 115, 124 or 131 are used, a more appropriate number of primers for positive individual identification would be 500-2,000.

If a panel of reagents from 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 nucleotide sequences described herein is used to generate a unique identification database for an individual, those same reagents can later be used to identify tissue from that individual. Using the unique identification database, positive identification of the individual, living or dead, can be made from extremely small tissue samples.

Use of Partial 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 Sequences in Forensic Biology

DNA-based identification techniques can also be used in forensic biology. To make such an identification, PCR technology can be used to amplify DNA sequences taken from very small biological samples such as tissues, e.g., hair or skin, or body fluids, e.g., blood, saliva, or semen found at a crime scene. The amplified sequence can then be compared to a standard, thereby allowing identification of the origin of the biological sample.

The sequences of the present invention can be used to provide polynucleotide reagents, e.g., PCR primers, targeted to specific loci in the human genome, which can enhance the reliability of DNA-based forensic identifications by, for example, providing another “identification marker” (i.e. another DNA sequence that is unique to a particular individual). As mentioned above, actual base sequence information can be used for identification as an accurate alternative to patterns formed by restriction enzyme generated fragments. Sequences targeted to noncoding regions of SEQ ID NO:1, 4, 7, 10, 13, 16, 53, 61, 67, 78, 88, 100, 113, 122 or 129 (e.g., fragments derived from the noncoding regions of SEQ ID NO:1, 4, 7, 10, 13, 16, 53, 61, 67, 78, 88, 100, 113, 122 or 129 having a length of at least 20 bases, preferably at least 30 bases) are particularly appropriate for this use.

The 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 nucleotide sequences described herein can further be used to provide polynucleotide reagents, e.g., labeled or labelable probes which can be used in, for example, an in situ hybridization technique, to identify a specific tissue. This can be very useful in cases where a forensic pathologist is presented with a tissue of unknown origin. Panels of such 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 probes can be used to identify tissue by species and/or by organ type.

In a similar fashion, these reagents, e.g., 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 primers or probes can be used to screen tissue culture for contamination (i.e. screen for the presence of a mixture of different types of cells in a culture).

Predictive Medicine

The present invention also pertains to the field of predictive medicine in which diagnostic assays, prognostic assays, and monitoring clinical trials are used for prognostic (predictive) purposes to thereby treat an individual.

Generally, the invention provides, a method of determining if a subject is at risk for a disorder related to a lesion in or the misexpression of a gene which encodes 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843.

Such disorders include, e.g., a disorder associated with the misexpression of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 gene; cellular proliferative and/or differentiative disorder, a brain, platelet, breast, colon, kidney (renal), lung, ovarian, prostate, hematopoeitic, pancreatic, skeletal muscle, skin (dermal), bone metabolism, immune, e.g., inflammatory, cardiovascular, endothelial cell, liver, viral diseases, pain, metabolic, neurological or CNS, erythroid or anemic disorder.

The method includes one or more of the following: detecting, in a tissue of the subject, the presence or absence of a mutation which affects the expression of the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 gene, or detecting the presence or absence of a mutation in a region which controls the expression of the gene, e.g., a mutation in the 5′ control region; detecting, in a tissue of the subject, the presence or absence of a mutation which alters the structure of the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 gene; detecting, in a tissue of the subject, the misexpression of the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 gene, at the mRNA level, e.g., detecting a non-wild type level of an mRNA; or detecting, in a tissue of the subject, the misexpression of the gene, at the protein level, e.g., detecting a non-wild type level of a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 polypeptide.

In preferred embodiments the method includes: ascertaining the existence of at least one of: a deletion of one or more nucleotides from the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 gene; an insertion of one or more nucleotides into the gene, a point mutation, e.g., a substitution of one or more nucleotides of the gene, a gross chromosomal rearrangement of the gene, e.g., a translocation, inversion, or deletion.

For example, detecting the genetic lesion can include: (i) providing a probe/primer including an oligonucleotide containing a region of nucleotide sequence which hybridizes to a sense or antisense sequence from SEQ ID NO:1, 4, 7, 10, 13, 16, 53, 61, 67, 78, 88, 100, 113, 122 or 129, or naturally occurring mutants thereof or 5′ or 3′ flanking sequences naturally associated with the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 gene; (ii) exposing the probe/primer to nucleic acid of the tissue; and detecting, by hybridization, e.g., in situ hybridization, of the probe/primer to the nucleic acid, the presence or absence of the genetic lesion.

In preferred embodiments detecting the misexpression includes ascertaining the existence of at least one of: an alteration in the level of a messenger RNA transcript of the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 gene; the presence of a non-wild type splicing pattern of a messenger RNA transcript of the gene; or a non-wild type level of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843.

Methods of the invention can be used prenatally or to determine if a subject's offspring will be at risk for a disorder.

In preferred embodiments the method includes determining the structure of a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 gene, an abnormal structure being indicative of risk for the disorder.

In preferred embodiments the method includes contacting a sample from the subject with an antibody to the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein or a nucleic acid, which hybridizes specifically with the gene. These and other embodiments are discussed below.

Diagnostic and Prognostic Assays

The presence, level, or absence of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein or nucleic acid in a biological sample can be evaluated by obtaining a biological sample from a test subject and contacting the biological sample with a compound or an agent capable of detecting 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein or nucleic acid (e.g., mRNA, genomic DNA) that encodes 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein such that the presence of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein or nucleic acid is detected in the biological sample. The term “biological sample” includes tissues, cells and biological fluids isolated from a subject, as well as tissues, cells and fluids present within a subject. A preferred biological sample is serum. The level of expression of the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 gene can be measured in a number of ways, including, but not limited to: measuring the mRNA encoded by the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 genes; measuring the amount of protein encoded by the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 genes; or measuring the activity of the protein encoded by the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 genes.

The level of mRNA corresponding to the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 gene in a cell can be determined both by in situ and by in vitro formats.

The isolated mRNA can be used in hybridization or amplification assays that include, but are not limited to, Southern or Northern analyses, polymerase chain reaction analyses and probe arrays. One preferred diagnostic method for the detection of mRNA levels involves contacting the isolated mRNA with a nucleic acid molecule (probe) that can hybridize to the mRNA encoded by the gene being detected. The nucleic acid probe can be, for example, a full-length 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 nucleic acid, such as the nucleic acid of SEQ ID NO:1, 4, 7, 10, 13, 16, 53, 61, 67, 78, 88, 100, 113, 122 or 129, or a portion thereof, such as an oligonucleotide of at least 7, 15, 30, 50, 100, 250 or 500 nucleotides in length and sufficient to specifically hybridize under stringent conditions to 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 mRNA or genomic DNA. Other suitable probes for use in the diagnostic assays are described herein.

In one format, mRNA (or cDNA) is immobilized on a surface and contacted with the probes, for example by running the isolated mRNA on an agarose gel and transferring the mRNA from the gel to a membrane, such as nitrocellulose. In an alternative formnat, the probes are immobilized on a surface and the mRNA (or CDNA) is contacted with the probes, for example, in a two-dimensional gene chip array. A skilled artisan can adapt known mRNA detection methods for use in detecting the level of mRNA encoded by the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 genes.

The level of mRNA in a sample that is encoded by one of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 can be evaluated with nucleic acid amplification, e.g., by rtPCR (Mullis (1987) U.S. Pat. No. 4,683,202), ligase chain reaction (Barany (1991) Proc. Natl. Acad. Sci. USA 88: 189-193), self sustained sequence replication (Guatelli et al., (1990) Proc. Natl. Acad. Sci. USA 87:1874-1878), transcriptional amplification system (Kwoh et al., (1989), Proc. Natl. Acad. Sci. USA 86:1173-1177), Q-Beta Replicase (Lizardi et al., (1988) Bio/Technology 6:1197), rolling circle replication (Lizardi et al., U.S. Pat. No. 5,854,033) or any other nucleic acid amplification method, followed by the detection of the amplified molecules using techniques known in the art. As used herein, amplification primers are defined as being a pair of nucleic acid molecules that can anneal to 5′ or 3′ regions of a gene (plus and minus strands, respectively, or vice-versa) and contain a short region in between. In general, amplification primers are from about 10 to 30 nucleotides in length and flank a region from about 50 to 200 nucleotides in length. Under appropriate conditions and with appropriate reagents, such primers permit the amplification of a nucleic acid molecule comprising the nucleotide sequence flanked by the primers.

For in situ methods, a cell or tissue sample can be prepared/processed and immobilized on a support, typically a glass slide, and then contacted with a probe that can hybridize to mRNA that encodes the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 gene being analyzed.

In another embodiment, the methods further contacting a control sample with a compound or agent capable of detecting 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 mRNA, or genomic DNA, and comparing the presence of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 mRNA or genomic DNA in the control sample with the presence of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 mRNA or genomic DNA in the test sample.

A variety of methods can be used to determine the level of protein encoded by 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843. In general, these methods include contacting an agent that selectively binds to the protein, such as an antibody with a sample, to evaluate the level of protein in the sample. In a preferred embodiment, the antibody bears a detectable label. Antibodies can be polyclonal, or more preferably, monoclonal. An intact antibody, or a fragment thereof (e.g., Fab or F(ab′) ₂) can be used. The term “labeled”, with regard to the probe or antibody, is intended to encompass direct labeling of the probe or antibody by coupling (i.e., physically linking) a detectable substance to the probe or antibody, as well as indirect labeling of the probe or antibody by reactivity with a detectable substance. Examples of detectable substances are provided herein.

The detection methods can be used to detect 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein in a biological sample in vitro as well as in vivo. In vitro techniques for detection of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein include enzyme linked immunosorbent assays (ELISAs), immunoprecipitations, immunofluorescence, enzyme immunoassay (EIA), radioimmunoassay (RIA), and Western blot analysis. In vivo techniques for detection of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein include introducing into a subject a labeled anti-26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 antibody. For example, the antibody can be labeled with a radioactive marker whose presence and location in a subject can be detected by standard imaging techniques.

In another embodiment, the methods further include contacting the control sample with a compound or agent capable of detecting 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein, and comparing the presence of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein in the control sample with the presence of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein in the test sample.

The invention also includes kits for detecting the presence of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 in a biological sample. For example, the kit can include a compound or agent capable of detecting 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein or mRNA in a biological sample; and a standard. The compound or agent can be packaged in a suitable container. The kit can further comprise instructions for using the kit to detect 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein or nucleic acid.

For antibody-based kits, the kit can include: (1) a first antibody (e.g., attached to a solid support) which binds to a polypeptide corresponding to a marker of the invention; and, optionally, (2) a second, different antibody which binds to either the polypeptide or the first antibody and is conjugated to a detectable agent.

For oligonucleotide-based kits, the kit can include: (1) an oligonucleotide, e.g., a detectably labeled oligonucleotide, which hybridizes to a nucleic acid sequence encoding a polypeptide corresponding to a marker of the invention or (2) a pair of primers useful for amplifying a nucleic acid molecule corresponding to a marker of the invention. The kit can also includes a buffering agent, a preservative, or a protein stabilizing agent. The kit can also includes components necessary for detecting the detectable agent (e.g., an enzyme or a substrate). The kit can also contain a control sample or a series of control samples which can be assayed and compared to the test sample contained. Each component of the kit can be enclosed within an individual container and all of the various containers can be within a single package, along with instructions for interpreting the results of the assays performed using the kit.

The diagnostic methods described herein can identify subjects having, or at risk of developing, a disease or disorder associated with misexpressed or aberrant or unwanted 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 expression or activity. As used herein, the term “unwanted” includes an unwanted phenomenon involved in a biological response such as pain or deregulated cell proliferation.

In one embodiment, a disease or disorder associated with aberrant or unwanted 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 expression or activity is identified. A test sample is obtained from a subject and 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein or nucleic acid (e.g., mRNA or genomic DNA) is evaluated, wherein the level, e.g., the presence or absence, of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein or nucleic acid is diagnostic for a subject having or at risk of developing a disease or disorder associated with aberrant or unwanted 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 expression or activity. As used herein, a “test sample” refers to a biological sample obtained from a subject of interest, including a biological fluid (e.g., serum), cell sample, or tissue.

The prognostic assays described herein can be used to determine whether a subject can be administered an agent (e.g., an agonist, antagonist, peptidomimetic, protein, peptide, nucleic acid, small molecule, or other drug candidate) to treat a disease or disorder associated with aberrant or unwanted 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 expression or activity. For example, such methods can be used to determine whether a subject can be effectively treated with an agent for a cellular proliferation or differentiation disorder, a brain, platelet, breast, colon, kidney (renal), lung, ovarian, prostate, hematopoeitic, pancreatic, skeletal muscle, skin (dermal), bone metabolism, immune, e.g., inflammatory, cardiovascular, endothelial cell, liver, viral diseases, pain, metabolic, neurological or CNS, erythroid or anemic disorder.

The methods of the invention can also be used to detect genetic alterations in a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 gene, thereby determining if a subject with the altered gene is at risk for a disorder characterized~by misregulation in 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein activity or nucleic acid expression, such as a a cellular proliferation or differentiation disorder, a brain, platelet, breast, colon, kidney (renal), lung, ovarian, prostate, hematopoeitic, pancreatic, skeletal muscle, skin (dermal), bone metabolism, immune, e.g., inflammatory, cardiovascular, endothelial cell, liver, viral diseases, pain, metabolic, neurological or CNS, erythroid or anemic disorder. In preferred embodiments, the methods include detecting, in a sample from the subject, the presence or absence of a genetic alteration characterized by at least one of an alteration affecting the integrity of a gene encoding a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843-protein, or the mis-expression of the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 gene. For example, such genetic alterations can be detected by ascertaining the existence of at least one of 1) a deletion of one or more nucleotides from a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 gene; 2) an addition of one or more nucleotides to a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 gene; 3) a substitution of one or more nucleotides of a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 gene, 4) a chromosomal rearrangement of a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 gene; 5) an alteration in the level of a messenger RNA transcript of a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 gene, 6) aberrant modification of a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 gene, such as of the methylation pattern of the genomic DNA, 7) the presence of a non-wild type splicing pattern of a messenger RNA transcript of a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 gene, 8) a non-wild type level of a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843-protein, 9) allelic loss of a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 gene, and 10) inappropriate post-translational modification of a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843-protein.

An alteration can be detected without a probe/primer in a polymerase chain reaction, such as anchor PCR or RACE PCR, or, alternatively, in a ligation chain reaction (LCR), the latter of which can be particularly useful for detecting point mutations in the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843-gene. This method can include the steps of collecting a sample of cells from a subject, isolating nucleic acid (e.g., genomic, mRNA or both) from the sample, contacting the nucleic acid sample with one or more primers which specifically hybridize to a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 gene under conditions such that hybridization and amplification of the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 gene (if present) occurs, and detecting the presence or absence of an amplification product, or detecting the size of the amplification product and comparing the length to a control sample. It is anticipated that PCR and/or LCR may be desirable to use as a preliminary amplification step in conjunction with any of the techniques used for detecting mutations described herein. Alternatively, other amplification methods described herein or known in the art can be used.

In another embodiment, mutations in a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 gene from a sample cell can be identified by detecting alterations in restriction enzyme cleavage patterns. For example, sample and control DNA is isolated, amplified (optionally), digested with one or more restriction endonucleases, and fragment length sizes are determined, e.g., by gel electrophoresis and compared. Differences in fragment length sizes between sample and control DNA indicates mutations in the sample DNA. Moreover, the use of sequence specific ribozymes (see, for example, U.S. Pat. No. 5,498,531) can be used to score for the presence of specific mutations by development or loss of a ribozyme cleavage site.

In other embodiments, genetic mutations in 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 can be identified by hybridizing a sample and control nucleic acids, e.g., DNA or RNA, two dimensional arrays, e.g., chip based arrays. Such arrays include a plurality of addresses, each of which is positionally distinguishable from the other. A different probe is located at each address of the plurality. The arrays can have a high density of addresses, e.g., can contain hundreds or thousands of oligonucleotides probes (Cronin et al. (1996) Human Mutation 7: 244-255; Kozal et al. (1996) Nature Medicine 2: 753-759). For example, genetic mutations in 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 can be identified in two dimensional arrays containing light-generated DNA probes as described in Cronin, M. T. et al. supra. Briefly, a first hybridization array of probes can be used to scan through long stretches of DNA in a sample and control to identify base changes between the sequences by making linear arrays of sequential overlapping probes. This step allows the identification of point mutations. This step is followed by a second hybridization array that allows the characterization of specific mutations by using smaller, specialized probe arrays complementary to all variants or mutations detected. Each mutation array is composed of parallel probe sets, one complementary to the wild-type gene and the other complementary to the mutant gene.

In yet another embodiment, any of a variety of sequencing reactions known in the art can be used to directly sequence the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 gene and detect mutations by comparing the sequence of the sample 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 with the corresponding wild-type (control) sequence. Automated sequencing procedures can be utilized when performing the diagnostic assays (Naeve et al. (1995) Biotechniques 19:448-53), including sequencing by mass spectrometry.

Other methods for detecting mutations in the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 gene include methods in which protection from cleavage agents is used to detect mismatched bases in RNA/RNA or RNA/DNA heteroduplexes (Myers et al. (1985) Science 230:1242; Cotton et al. (1988) Proc. Natl Acad Sci USA 85:4397; Saleeba et al. (1992) Methods Enzymol. 217:286-295).

In still another embodiment, the mismatch cleavage reaction employs one or more proteins that recognize mismatched base pairs in double-stranded DNA (so called “DNA mismatch repair” enzymes) in defined systems for detecting and mapping point mutations in 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 cDNAs obtained from samples of cells. For example, the mutY enzyme of E. coli cleaves A at G/A mismatches and the thymidine DNA glycosylase from HeLa cells cleaves T at G/T mismatches (Hsu et al. (1994) Carcinogenesis 15:1657-1662; U.S. Pat. No. 5,459,039).

In other embodiments, alterations in electrophoretic mobility will be used to identify mutations in 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 genes. For example, single strand conformation polymorphism (SSCP) can be used to detect differences in electrophoretic mobility between mutant and wild type nucleic acids (Orita et al. (1989) Proc Natl. Acad. Sci USA: 86:2766, see also Cotton (1993) Mutat. Res. 285:125-144; and Hayashi (1992) Genet. Anal. Tech. Appl. 9:73-79). Single-stranded DNA fragments of sample and control 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 nucleic acids will be denatured and allowed to renature. The secondary structure of single-stranded nucleic acids varies according to sequence, the resulting alteration in electrophoretic mobility enables the detection of even a single base change. The DNA fragments can be labeled or detected with labeled probes. The sensitivity of the assay can be enhanced by using RNA (rather than DNA), in which the secondary structure is more sensitive to a change in sequence. In a preferred embodiment, the subject method utilizes heteroduplex analysis to separate double stranded heteroduplex molecules on the basis of changes in electrophoretic mobility (Keen et al. (1991) Trends Genet 7:5).

In yet another embodiment, the movement of mutant or wild-type fragments in polyacrylamide gels containing a gradient of denaturant is assayed using denaturing gradient gel electrophoresis (DGGE) (Myers et al. (1985) Nature 313:495). When DGGE is used as the method of analysis, DNA will be modified to insure that it does not completely denature, for example by adding a GC clamp of approximately 40 bp of high-melting GC-rich DNA by PCR. In a further embodiment, a temperature gradient is used in place of a denaturing gradient to identify differences in the mobility of control and sample DNA (Rosenbaum and Reissner (1987) Biophys Chem 265:12753).

Examples of other techniques for detecting point mutations include, but are not limited to, selective oligonucleotide hybridization, selective amplification, or selective primer extension (Saiki et al. (1986) Nature 324:163); Saiki et al. (1989) Proc. Natl Acad. Sci USA 86:6230).

Alternatively, allele specific amplification technology which depends on selective PCR amplification can be used in conjunction with the instant invention. Oligonucleotides used as primers for specific amplification can carry the mutation of interest in the center of the molecule (so that amplification depends on differential hybridization) (Gibbs et al. (1989) Nucleic Acids Res. 17:2437-2448) or at the extreme 3′end of one primer where, under appropriate conditions, mismatch can prevent, or reduce polymerase extension (Prossner (1993) Tibtech 11:238). In addition it may be desirable to introduce a novel restriction site in the region of the mutation to create cleavage-based detection (Gasparini et al. (1992) Mol. Cell Probes 6: 1). It is anticipated that in certain embodiments amplification can also be performed using Taq ligase for amplification (Barany (1991) Proc. Natl. Acad. Sci USA 88:189-93). In such cases, ligation will occur only if there is a perfect match at the 3′ end of the 5′ sequence making it possible to detect the presence of a known mutation at a specific site by looking for the presence or absence of amplification.

The methods described herein can be performed, for example, by utilizing pre-packaged diagnostic kits comprising at least one probe nucleic acid or antibody reagent described herein, which can be conveniently used, e.g., in clinical settings to diagnose patients exhibiting symptoms or family history of a disease or illness involving a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 gene.

Use of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 Molecules as Surrogate Markers

The 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 molecules of the invention are also useful as markers of disorders or disease states, as markers for precursors of disease states, as markers for predisposition of disease states, as markers of drug activity, or as markers of the pharmacogenomic profile of a subject. Using the methods described herein, the presence, absence and/or quantity of the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 molecules of the invention can be detected, and can be correlated with one or more biological states in vivo. For example, the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 molecules of the invention can serve as surrogate markers for one or more disorders or disease states or for conditions leading up to disease states. As used herein, a “surrogate marker” is an objective biochemical marker which correlates with the absence or presence of a disease or disorder, or with the progression of a disease or disorder (e.g., with the presence or absence of a tumor). The presence or quantity of such markers is independent of the disease. Therefore, these markers can serve to indicate whether a particular course of treatment is effective in lessening a disease state or disorder. Surrogate markers are of particular use when the presence or extent of a disease state or disorder is difficult to assess through standard methodologies (e.g., early stage tumors), or when an assessment of disease progression is desired before a potentially dangerous clinical endpoint is reached (e.g., an assessment of cardiovascular disease can be made using cholesterol levels as a surrogate marker, and an analysis of HIV infection can be made using HIV RNA levels as a surrogate marker, well in advance of the undesirable clinical outcomes of myocardial infarction or fully-developed AIDS). Examples of the use of surrogate markers in the art include: Koomen et al. (2000) J. Mass. Spectrom. 35: 258-264; and James (1994) AIDS Treatment News Archive 209.

The 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 molecules of the invention are also useful as pharmacodynamic markers. As used herein, a “pharmacodynamic marker” is an objective biochemical marker which correlates specifically with drug effects. The presence or quantity of a pharmacodynamic marker is not related to the disease state or disorder for which the drug is being administered; therefore, the presence or quantity of the marker is indicative of the presence or activity of theadrug in a subject. For example, a pharmacodynamic marker can be indicative of the concentration of the drug in a biological tissue, in that the marker is either expressed or transcribed or not expressed or transcribed in that tissue in relationship to the level of the drug. In this fashion, the distribution or uptake of the drug can be monitored by the pharmacodynamic marker. Similarly, the presence or quantity of the pharmacodynamic marker can be related to the presence or quantity of the metabolic product of a drug, such that the presence or quantity of the marker is indicative of the relative breakdown rate of the drug in vivo. Pharmacodynamic markers are of particular use in increasing the sensitivity of detection of drug effects, particularly when the drug is administered in low doses. Since even a small amount of a drug can be sufficient to activate multiple rounds of marker (e.g., a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 marker) transcription or expression, the amplified marker can be in a quantity which is more readily detectable than the drug itself. Also, the marker can be more easily detected due to the nature of the marker itself; for example, using the methods described herein, anti-26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 antibodies can be employed in an immune-based detection system for a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein marker, or 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843-specific radiolabeled probes can be used to detect a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 mRNA marker. Furthermore, the use of a pharmacodynamic marker can offer mechanism-based prediction of risk due to drug treatment beyond the range of possible direct observations. Examples of the use of pharmacodynamic markers in the art include: Matsuda et al. U.S. Pat. No. 6,033,862; Hattis et al. (1991) Env. Health Perspect. 90: 229-238; Schentag (1999) Am. J. Health-Syst. Pharm. 56 Suppl. 3: S21-S24; and Nicolau (1999) Am. J. Health-Syst. Pharm. 56 Suppl. 3: S16-S20.

The 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 molecules of the invention are also useful as pharmacogenomic markers. As used herein, a “pharmacogenomic marker” is an objective biochemical marker which correlates with a specific clinical drug response or susceptibility in a subject (see, e.g., McLeod et al. (1999) Eur. J. Cancer 35:1650-1652). The presence or quantity of the pharmacogenomic marker is related to the predicted response of the subject to a specific drug or class of drugs prior to administration of the drug. By assessing the presence or quantity of one or more pharmacogenomic markers in a subject, a drug therapy which is most appropriate for the subject, or which is predicted to have a greater degree of success, can be selected. For example, based on the presence or quantity of RNA, or protein (e.g., 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein or RNA) for specific tumor markers in a subject, a drug or course of treatment can be selected that is optimized for the treatment of the specific tumor likely to be present in the subject. Similarly, the presence or absence of a specific sequence mutation in 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 DNA can correlate with a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 drug response. The use of pharmacogenomic markers therefore permits the application of the most appropriate treatment for each subject without having to administer the therapy.

Pharmaceutical Compositions

The nucleic acid and polypeptides, fragments thereof, as well as anti-26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 antibodies (also referred to herein as “active compounds”) of the invention can be incorporated into pharmaceutical compositions. Such compositions typically include the nucleic acid molecule, protein, or antibody and a pharmaceutically acceptable carrier. As used herein the language “pharmaceutically acceptable carrier” includes solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like, compatible with pharmaceutical administration. Supplementary active compounds can also be incorporated into the compositions.

A pharmaceutical composition is formulated to be compatible with its intended route of administration. Examples of routes of administration include parenteral, e.g., intravenous, intradermal, subcutaneous, oral (e.g., inhalation), transdermal (topical), transmucosal, and rectal administration. Solutions or suspensions used for parenteral, intradermal, or subcutaneous application can include the following components: a sterile diluent such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerine, propylene glycol or other synthetic solvents; antibacterial agents such as benzyl alcohol or methyl parabens; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid; buffers such as acetates, citrates or phosphates and agents for the adjustment of tonicity such as sodium chloride or dextrose. pH can be adjusted with acids or bases, such as hydrochloric acid or sodium hydroxide. The parenteral preparation can be enclosed in ampoules, disposable syringes or multiple dose vials made of glass or plastic.

Pharmaceutical compositions suitable for injectable use include sterile aqueous solutions (where water soluble) or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersion. For intravenous administration, suitable carriers include physiological saline, bacteriostatic water, Cremophor EL™ (BASF, Parsippany, N.J.) or phosphate buffered saline (PBS). In all cases, the composition must be sterile and should be fluid to the extent that easy syringability exists. It should be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms such as bacteria and fungi. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyetheylene glycol, and the like), and suitable mixtures thereof. The proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants. Prevention of the action of microorganisms can be achieved by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars, polyalcohols such as manitol, sorbitol, sodium chloride in the composition. Prolonged absorption of the injectable compositions can be brought about by including in the composition an agent which delays absorption, for example, aluminum monostearate and gelatin.

Sterile injectable solutions can be prepared by incorporating the active compound in the required amount in an appropriate solvent with one or a combination of ingredients enumerated above, as required, followed by filtered sterilization. Generally, dispersions are prepared by incorporating the active compound into a sterile vehicle which contains a basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum drying and freeze-drying which yields a powder of the active ingredient plus any additional desired ingredient from a previously sterile-filtered solution thereof.

Oral compositions generally include an inert diluent or an edible carrier. For the purpose of oral therapeutic administration, the active compound can be incorporated with excipients and used in the form of tablets, troches, or capsules, e.g., gelatin capsules. Oral compositions can also be prepared using a fluid carrier for use as a mouthwash. Pharmaceutically compatible binding agents, and/or adjuvant materials can be included as part of the composition. The tablets, pills, capsules, troches and the like can contain any of the following ingredients, or compounds of a similar nature: a binder such as microcrystalline cellulose, gum tragacanth or gelatin; an excipient such as starch or lactose, a disintegrating agent such as alginic acid, Primogel, or corn starch; a lubricant such as magnesium stearate or Sterotes; a glidant such as colloidal silicon dioxide; a sweetening agent such as sucrose or saccharin; or a flavoring agent such as peppermint, methyl salicylate, or orange flavoring.

For administration by inhalation, the compounds are delivered in the form of an aerosol spray from pressured container or dispenser which contains a suitable propellant, e.g., a gas such as carbon dioxide, or a nebulizer.

Systemic administration can also be by transmucosal or transdermal means. For transmucosal or transdermal administration, penetrants appropriate to the barrier to be permeated are used in the formulation. Such penetrants are generally known in the art, and include, for example, for transmucosal administration, detergents, bile salts, and fusidic acid derivatives. Transmucosal administration can be accomplished through the use of nasal sprays or suppositories. For transdermal administration, the active compounds are formulated into ointments, salves, gels, or creams as generally known in the art.

The compounds can also be prepared in the form of suppositories (e.g., with conventional suppository bases such as cocoa butter and other glycerides) or retention enemas for rectal delivery.

In one embodiment, the active compounds are prepared with carriers that will protect the compound against rapid elimination from the body, such as a controlled release formulation, including implants and microencapsulated delivery systems. Biodegradable, biocompatible polymers can be used, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid. Methods for preparation of such formulations will be apparent to those skilled in the art. The materials can also be obtained commercially from Alza Corporation and Nova Pharmaceuticals, Inc. Liposomal suspensions (including liposomes targeted to infected cells with monoclonal antibodies to viral antigens) can also be used as pharmaceutically acceptable carriers. These can be prepared according to methods known to those skilled in the art, for example, as described in U.S. Pat. No. 4,522,811.

It is advantageous to formulate oral or parenteral compositions in dosage unit form for ease of administration and uniformity of dosage. Dosage unit form as used herein refers to physically discrete units suited as unitary dosages for the subject to be treated; each unit containing a predetermined quantity of active compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier.

Toxicity and therapeutic efficacy of such compounds can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, e.g., for determining the LD ₅₀(the dose lethal to 50% of the population) and the ED₅₀(the dose therapeutically effective in 50% of the population). The dose ratio between toxic and therapeutic effects is the therapeutic index and it can be expressed as the ratio LD₅₀/ ED₅₀-Compounds which exhibit high therapeutic indices are preferred. While compounds that exhibit toxic side effects can be used, care should be taken to design a delivery system that targets such compounds to the site of affected tissue in order to minimize potential damage to uninfected cells and, thereby, reduce side effects.

The data obtained from the cell culture assays and animal studies can be used in formulating a range of dosage for use in humans. The dosage of such compounds lies preferably within a range of circulating concentrations that include the ED ₅₀with little or no toxicity. The dosage can vary within this range depending upon the dosage form employed and the route of administration utilized. For any compound used in the method of the invention, the therapeutically effective dose can be estimated initially from cell culture assays. A dose can be formulated in animal models to achieve a circulating plasma concentration range that includes the IC₅₀(i.e., the concentration of the test compound which achieves a half-maximal inhibition of symptoms) as determined in cell culture. Such information can be used to more accurately determine useful doses in humans. Levels in plasma can be measured, for example, by high performance liquid chromatography.

As defined herein, a therapeutically effective amount of protein or polypeptide (i.e., an effective dosage) ranges from about 0.001 to 30 mg/kg body weight, preferably about 0.01 to 25 mg/kg body weight, more preferably about 0.1 to 20 mg/kg body weight, and even more preferably about 1 to 10 mg/kg, 2 to 9 mg/kg, 3 to 8 mg/kg, 4 to 7 mg/kg, or 5 to 6 mg/kg body weight. The protein or polypeptide can be administered one time per week for between about 1 to 10 weeks, preferably between 2 to 8 weeks, more preferably between about 3 to 7 weeks, and even more preferably for about 4, 5, or 6 weeks. The skilled artisan will appreciate that certain factors can influence the dosage and timing required to effectively treat a subject, including but not limited to the severity of the disease or disorder, previous treatments, the general health and/or age of the subject, and other diseases present. Moreover, treatment of a subject with a therapeutically effective amount of a protein, polypeptide, or antibody, unconjugated or conjugated as described herein, can include a single treatment or, preferably, can include a series of treatments.

For antibodies, the preferred dosage is 0.1 mg/kg of body weight (generally 10 mg/kg to 20 mg/kg). If the antibody is to act in the brain, a dosage of 50 mg/kg to 100 mg/kg is usually appropriate. Generally, partially human antibodies and fully human antibodies have a longer half-life within the human body than other antibodies. Accordingly, lower dosages and less frequent administration is often possible. Modifications such as lipidation can be used to stabilize antibodies and to enhance uptake and tissue penetration (e.g., into the brain). A method for lipidation of antibodies is described by Cruikshank et al. ((1997) J. Acquired Immune Deficiency Syndromes and Human Retrovirology 14:193).

The present invention encompasses agents which modulate expression or activity. An agent can, for example, be a small molecule. For example, such small molecules include, but are not limited to, peptides, peptidomimetics (e.g., peptoids), amino acids, amino acid analogs, polynucleotides, polynucleotide analogs, nucleotides, nucleotide analogs, organic or inorganic compounds (i.e., including heteroorganic and organometallic compounds) having a molecular weight less than about 10,000 grams per mole, organic or inorganic compounds having a molecular weight less than about 5,000 grams per mole, organic or inorganic compounds having a molecular weight less than about 1,000 grams per mole, organic or inorganic compounds having a molecular weight less than about 500 grams per mole, and salts, esters, and other pharmaceutically acceptable forms of such compounds.

Exemplary doses include milligram or microgram amounts of the small molecule per kilogram of subject or sample weight (e.g., about 1 microgram per kilogram to about 500 milligrams per kilogram, about 100 micrograms per kilogram to about 5 milligrams per kilogram, or about 1 microgram per kilogram to about 50 micrograms per kilogram. It is furthermore understood that appropriate doses of a small molecule depend upon the potency of the small molecule with respect to the expression or activity to be modulated. When one or more of these small molecules is to be administered to an animal (e.g., a human) in order to modulate expression or activity of a polypeptide or nucleic acid of the invention, a physician, veterinarian, or researcher can, for example, prescribe a relatively low dose at first, subsequently increasing the dose until an appropriate response is obtained. In addition, it is understood that the specific dose level for any particular animal subject will depend upon a variety of factors including the activity of the specific compound employed, the age, body weight, general health, gender, and diet of the subject, the time of administration, the route of administration, the rate of excretion, any drug combination, and the degree of expression or activity to be modulated.

The nucleic acid molecules of the invention can be inserted into vectors and used as gene therapy vectors. Gene therapy vectors can be delivered to a subject by, for example, intravenous injection, local administration (see U.S. Pat. No. 5,328,470) or by stereotactic injection (see e.g., Chen et al. (1994) Proc. Natl. Acad. Sci. USA 91:3054-3057). The pharmaceutical preparation of the gene therapy vector can include the gene therapy vector in an acceptable diluent, or can comprise a slow release matrix in which the gene delivery vehicle is imbedded. Alternatively, where the complete gene delivery vector can be produced intact from recombinant cells, e.g., retroviral vectors, the pharmaceutical preparation can include one or more cells which produce the gene delivery system.

The pharmaceutical compositions can be included in a container, pack, or dispenser together with instructions for administration.

Methods of Treatment:

The present invention provides for both prophylactic and therapeutic methods of treating a subject at risk of (or susceptible to) a disorder or having a disorder associated with aberrant or unwanted 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 expression or activity. As used herein, the term “treatment” is defined as the application or administration of a therapeutic agent to a patient, or application or administration of a therapeutic agent to an isolated tissue or cell line from a patient, who has a disease, a symptom of disease or a predisposition toward a disease, with the purpose to cure, heal, alleviate, relieve, alter, remedy, ameliorate, improve or affect the disease, the symptoms of disease or the predisposition toward disease. A therapeutic agent includes, but is not limited to, small molecules, peptides, antibodies, ribozymes and antisense oligonucleotides.

With regards to both prophylactic and therapeutic methods of treatment, such treatments can be specifically tailored or modified, based on knowledge obtained from the field of pharmacogenomics. “Pharmacogenomics”, as used herein, refers to the application of genomics technologies such as gene sequencing, statistical genetics, and gene expression analysis to drugs in clinical development and on the market. More specifically, the term refers the study of how a patient's genes determine his or her response to a drug (e.g., a patient's “drug response phenotype”, or “drug response genotype”.) Thus, another aspect of the invention provides methods for tailoring an individual's prophylactic or therapeutic treatment with either the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 molecules of the present invention or 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 modulators according to that individual's drug response genotype. Pharmacogenomics allows a clinician or physician to target prophylactic or therapeutic treatments to patients who will most benefit from the treatment and to avoid treatment of patients who will experience toxic drug-related side effects.

In one aspect, the invention provides a method for preventing in a subject, a disease or condition associated with an aberrant or unwanted 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 expression or activity, by administering to the subject a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 or an agent which modulates 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 expression or at least one 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 activity. Subjects at risk for a disease which is caused or contributed to by aberrant or unwanted 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 expression or activity can be identified by, for example, any or a combination of diagnostic or prognostic assays as described herein. Administration of a prophylactic agent can occur prior to the manifestation of symptoms characteristic of the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 aberrance, such that a disease or disorder is prevented or, alternatively, delayed in its progression. Depending on the type of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 aberrance, for example, a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843, 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 agonist or 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 antagonist agent can be used for treating the subject. The appropriate agent can be determined based on screening assays described herein.

It is possible that some 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 disorders can be caused, at least in part, by an abnormal level of gene product, or by the presence of a gene product exhibiting abnormal activity. As such, the reduction in the level and/or activity of such gene products would bring about the amelioration of disorder symptoms.

The 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 molecules can act as novel diagnostic targets and therapeutic agents for controlling one or more of a cellular proliferation and/or differentiation disorder, a brain, platelet, breast, colon, kidney (renal), lung, ovarian, prostate, hematopoeitic, pancreatic, skeletal muscle, skin (derrnal), bone metabolism, immune, e.g., inflammatory, cardiovascular, endothelial cell, liver, viral diseases, pain, metabolic, neurological or CNS, erythroid or anemic disorder, all of which are described above.

As discussed, successful treatment of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 disorders can be brought about by techniques that serve to inhibit the expression or activity of target gene products. For example, compounds, e.g., an agent identified using an assays described above, that proves to exhibit negative modulatory activity, can be used in accordance with the invention to prevent and/or ameliorate symptoms of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 disorders. Such molecules can include, but are not limited to peptides, phosphopeptides, small organic or inorganic molecules, or antibodies (including, for example, polyclonal, monoclonal, humanized, human, anti-idiotypic, chimeric or single chain antibodies, and Fab, F(ab′) ₂and Fab expression library fragments, scFV molecules, and epitope-binding fragments thereof).

Further, antisense and ribozyme molecules that inhibit expression of the target gene can also be used in accordance with the invention to reduce the level of target gene expression, thus effectively reducing the level of target gene activity. Still further, triple helix molecules can be utilized in reducing the level of target gene activity. Antisense, ribozyme and triple helix molecules are discussed above.

It is possible that the use of antisense, ribozyme, and/or triple helix molecules to reduce or inhibit mutant gene expression can also reduce or inhibit the transcription (triple helix) and/or translation (antisense, ribozyme) of mRNA produced by normal target gene alleles, such that the concentration of normal target gene product present can be lower than is necessary for a normal phenotype. In such cases, nucleic acid molecules that encode and express target gene polypeptides exhibiting normal target gene activity can be introduced into cells via gene therapy method. Alternatively, in instances in that the target gene encodes an extracellular protein, it can be preferable to co-administer normal target gene protein into the cell or tissue in order to maintain the requisite level of cellular or tissue target gene activity.

Another method by which nucleic acid molecules can be utilized in treating or preventing a disease characterized by 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 expression is through the use of aptamer molecules specific for 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein. Aptamers are nucleic acid molecules having a tertiary structure which permits them to specifically or selectively bind to protein ligands (see, e.g., Osborne et al. (1997) Curr. Opin. Chem Biol. 1: 5-9; and Patel (1997) Curr Opin Chem Biol 1:32-46). Since nucleic acid molecules can in many cases be more conveniently introduced into target cells than therapeutic protein molecules can be, aptamers offer a method by which 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein activity can be specifically decreased without the introduction of drugs or other molecules which can have pluripotent effects.

Antibodies can be generated that are both specific for target gene product and that reduce target gene product activity. Such antibodies can, therefore, by administered in instances whereby negative modulatory techniques are appropriate for the treatment of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 disorders. For a description of antibodies, see the Antibody section above.

In circumstances wherein injection of an animal or a human subject with a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein or epitope for stimulating antibody production is harmful to the subject, it is possible to generate an immune response against 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 through the use of anti-idiotypic antibodies (see, for example, Herlyn (1999) Ann Med 31:66-78; and Bhattacharya-Chatteijee and Foon (1998) Cancer Treat Res. 94:51-68). If an anti-idiotypic antibody is introduced into a mammal or human subject, it should stimulate the production of anti-anti-idiotypic antibodies, which should be specific to the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein.

Vaccines directed to a disease characterized by 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 expression can also be generated in this fashion.

In instances where the target antigen is intracellular and whole antibodies are used, internalizing antibodies can be preferred. Lipofectin or liposomes can be used to deliver the antibody or a fragment of the Fab region that binds to the target antigen into cells. Where fragments of the antibody are used, the smallest inhibitory fragment that binds to the target antigen is preferred. For example, peptides having an amino acid sequence corresponding to the Fv region of the antibody can be used. Alternatively, single chain neutralizing antibodies that bind to intracellular target antigens can also be administered. Such single chain antibodies can be administered, for example, by expressing nucleotide sequences encoding single-chain antibodies within the target cell population (see e.g., Marasco et al. (1993) Proc. Natl. Acad. Sci. USA 90:7889-7893).

The identified compounds that inhibit target gene expression, synthesis and/or activity can be administered to a patient at therapeutically effective doses to prevent, treat or ameliorate 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 disorders. A therapeutically effective dose refers to that amount of the compound sufficient to result in amelioration of symptoms of the disorders. Toxicity and therapeutic efficacy of such compounds can be determined by standard pharmaceutical procedures as described above.

The data obtained from the cell culture assays and animal studies can be used in formulating a range of dosage for use in humans. The dosage of such compounds lies preferably within a range of circulating concentrations that include the ED ₅₀with little or no toxicity. The dosage can vary within this range depending upon the dosage form employed and the route of administration utilized. For any compound used in the method of the invention, the therapeutically effective dose can be estimated initially from cell culture assays. A dose can be formulated in animal models to achieve a circulating plasma concentration range that includes the IC₅₀(i.e., the concentration of the test compound that achieves a half-maximal inhibition of symptoms) as determined in cell culture. Such information can be used to more accurately determine useful doses in humans. Levels in plasma can be measured, for example, by high performance liquid chromatography.

Another example of determination of effective dose for an individual is the ability to directly assay levels of “free” and “bound” compound in the serum of the test subject. Such assays can utilize antibody mimics and/or “biosensors” that have been created through molecular imprinting techniques. The compound which is able to modulate 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 activity is used as a template, or “imprinting molecule”, to spatially organize polymerizable monomers prior to their polymerization with catalytic reagents. The subsequent removal of the imprinted molecule leaves a polymer matrix which contains a repeated “negative image” of the compound and is able to selectively rebind the molecule under biological assay conditions. A detailed review of this technique can be seen in Ansell et al (1996) Current Opinion in Biotechnology 7:89-94 and in Shea (1994) Trends in Polymer Science 2:166-173. Such “imprinted” affinity matrixes are amenable to ligand-binding assays, whereby the immobilized monoclonal antibody component is replaced by an appropriately imprinted matrix. An example of the use of such matrixes in this way can be seen in Vlatakis et al (1993) Nature 361:645-647. Through the use of isotope-labeling, the “free” concentration of compound which modulates the expression or activity of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 can be readily monitored and used in calculations of IC₅₀.

Such “imprinted” affinity matrixes can also be designed to include fluorescent groups whose photon-emitting properties measurably change upon local and selective binding of target compound. These changes can be readily assayed in real time using appropriate fiberoptic devices, in turn allowing the dose in a test subject to be quickly optimized based on its individual IC ₅₀. An rudimentary example of such a “biosensor” is discussed in Kriz et al (1995) Analytical Chemistry 67:2142-2144.

Another aspect of the invention pertains to methods of modulating 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 expression or activity for therapeutic purposes. Accordingly, in an exemplary embodiment, the modulatory method of the invention involves contacting a cell with a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 or agent that modulates one or more of the activities of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein activity associated with the cell. An agent that modulates 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein activity can be an agent as described herein, such as a nucleic acid or a protein, a naturally-occurring target molecule of a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein (e.g., a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 substrate or receptor), a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 antibody, a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 agonist or antagonist, a peptidomimetic of a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 agonist or antagonist, or other small molecule.

In one embodiment, the agent stimulates one or 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 activities. Examples of such stimulatory agents include active 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein and a nucleic acid molecule encoding 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843. In another embodiment, the agent inhibits one or more 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 activities. Examples of such inhibitory agents include antisense 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 nucleic acid molecules, anti-26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 antibodies, and 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 inhibitors. These modulatory methods can be performed in vitro (e.g., by culturing the cell with the agent) or, alternatively, in vivo (e.g., by administering the agent to a subject). As such, the present invention provides methods of treating an individual afflicted with a disease or disorder characterized by aberrant or unwanted expression or activity of a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein or nucleic acid molecule. In one embodiment, the method involves administering an agent (e.g., an agent identified by a screening assay described herein), or combination of agents that modulates (e.g., up regulates or down regulates) 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 expression or activity. In another embodiment, the method involves administering a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein or nucleic acid molecule as therapy to compensate for reduced, aberrant, or unwanted 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 expression or activity.

Stimulation of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 activity is desirable in situations in which 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 is abnormally downregulated and/or in which increased 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 activity is likely to have a beneficial effect. For example, stimulation of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 activity is desirable in situations in which a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 is downregulated and/or in which increased 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 activity is likely to have a beneficial effect. Likewise, inhibition of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 activity is desirable in situations in which 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 is abnormally upregulated and/or in which decreased 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 activity is likely to have a beneficial effect.

Pharmacogenomics

The 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 molecules of the present invention, as well as agents, or modulators which have a stimulatory or inhibitory effect on 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 activity (e.g., 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 gene expression) as identified by a screening assay described herein can be administered to individuals to treat (prophylactically or therapeutically) 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843-associated disorders (e.g., aberrant or deficient arginine methyltransferase activity, glycosyltransferase activity, gamma-glutamyltraspeptidase activity, phosphoribosylglycinamide transferase activity, acyltransferase activity, acyl-CoA dehydrogenase activity, fatty acid amide hydrolase activity, aminotransferase activity, zinc carboxypeptidase activity, protein kinase activity, DEAD helicase activity, short-chain dehydrogenase/reductase activity or phosphatase activity) associated with aberrant or unwanted 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 activity.

In conjunction with such treatment, pharmacogenomics (i.e., the study of the relationship between an individual's genotype and that individual's response to a foreign compound or drug) can be considered. Differences in metabolism of therapeutics can lead to severe toxicity or therapeutic failure by altering the relation between dose and blood concentration of the pharmacologically active drug. Thus, a physician or clinician can consider applying knowledge obtained in relevant pharmacogenomics studies in determining whether to administer a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 molecule or 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 modulator as well as tailoring the dosage and/or therapeutic regimen of treatment with a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 molecule or 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 modulator.

Pharmacogenomics deals with clinically significant hereditary variations in the response to drugs due to altered drug disposition and abnormal action in affected persons. See, for example, Eichelbaum et al. (1996) Clin. Exp. Pharmacol. Physiol. 23:983-985 and Linder et al. (1997) Clin. Chem. 43:254-266. In general, two types of pharmacogenetic conditions can be differentiated. Genetic conditions transmitted as a single factor altering the way drugs act on the body (altered drug action) or genetic conditions transmitted as single factors altering the way the body acts on drugs (altered drug metabolism). These pharmacogenetic conditions can occur either as rare genetic defects or as naturally-occurring polymorphisms. For example, glucose-6-phosphate dehydrogenase deficiency (G6PD) is a common inherited enzymopathy in which the main clinical complication is haemolysis after ingestion of oxidant drugs (anti-malarials, sulfonamides, analgesics, nitrofurans) and consumption of fava beans.

One pharmacogenomics approach to identifying genes that predict drug response, known as “a genome-wide association”, relies primarily on a high-resolution map of the human genome consisting of already known gene-related markers (e.g., a “bi-allelic” gene marker map which consists of 60,000-100,000 polymorphic or variable sites on the human genome, each of which has two variants.) Such a high-resolution genetic map can be compared to a map of the genome of each of a statistically significant number of patients taking part in a Phase II/III drug trial to identify markers associated with a particular observed drug response or side effect. Alternatively, such a high resolution map can be generated from a combination of some ten-million known single nucleotide polymorphisms (SNPs) in the human genome. As used herein, a “SNP” is a common alteration that occurs in a single nucleotide base in a stretch of DNA. For example, a SNP can occur once per every 1000 bases of DNA. A SNP can be involved in a disease process, however, the vast majority can not be disease-associated. Given a genetic map based on the occurrence of such SNPs, individuals can be grouped into genetic categories depending on a particular pattern of SNPs in their individual genome. In such a manner, treatment regimens can be tailored to groups of genetically similar individuals, taking into account traits that can be common among such genetically similar individuals.

Alternatively, a method termed the “candidate gene approach”, can be utilized to identify genes that predict drug response. According to this method, if a gene that encodes a drug's target is known (e.g., a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein of the present invention), all common variants of that gene can be fairly easily identified in the population and it can be determined if having one version of the gene versus another is associated with a particular drug response.

Alternatively, a method termed the “gene expression profiling”, can be utilized to identify genes that predict drug response. For example, the gene expression of an animal dosed with a drug (e.g., a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 molecule or 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 modulator of the present invention) can give an indication whether gene pathways related to toxicity have been turned on.

Information generated from more than one of the above pharmacogenomics approaches can be used to determine appropriate dosage and treatment regimens for prophylactic or therapeutic treatment of an individual. This knowledge, when applied to dosing or drug selection, can avoid adverse reactions or therapeutic failure and thus enhance therapeutic or prophylactic efficiency when treating a subject with a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 molecule or 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 modulator, such as a modulator identified by one of the exemplary screening assays described herein.

The present invention further provides methods for identifying new agents, or combinations, that are based on identifying agents that modulate the activity of one or more of the gene products encoded by one or more of the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 genes of the present invention, wherein these products can be associated with resistance of the cells to a therapeutic agent. Specifically, the activity of the proteins encoded by the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 genes of the present invention can be used as a basis for identifying agents for overcoming agent resistance. By blocking the activity of one or more of the resistance proteins, target cells, e.g., human cells, will become sensitive to treatment with an agent to which the unmodified target cells were resistant.

Monitoring the influence of agents (e.g., drugs) on the expression or activity of a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein can be applied in clinical trials. For example, the effectiveness of an agent determined by a screening assay as described herein to increase 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 gene expression, protein levels, or upregulate 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 activity, can be monitored in clinical trials of subjects exhibiting decreased 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 gene expression, protein levels, or downregulated 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 activity. Alternatively, the effectiveness of an agent determined by a screening assay to decrease 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 gene expression, protein levels, or downregulate 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 activity, can be monitored in clinical trials of subjects exhibiting increased 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 gene expression, protein levels, or upregulated 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 activity. In such clinical trials, the expression or activity of a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 gene, and preferably, other genes that have been implicated in, for example, a protein kinase-associated or another 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843-associated disorder can be used as a “read out” or markers of the phenotype of a particular cell.

Other Embodiments

In another aspect, the invention features a method of analyzing a plurality of capture probes. The method is useful, e.g., to analyze gene expression. The method includes: providing a two dimensional array having a plurality of addresses, each address of the plurality being positionally distinguishable from each other address of the plurality, and each address of the plurality having a unique capture probe, e.g., a nucleic acid or peptide sequence, wherein the capture probes are from a cell or subject which expresses 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 or from a cell or subject in which a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 mediated response has been elicited; contacting the array with a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 nucleic acid (preferably purified), a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 polypeptide (preferably purified), or an anti-26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 antibody, and thereby evaluating the plurality of capture probes. Binding, e.g., in the case of a nucleic acid, hybridization with a capture probe at an address of the plurality, is detected, e.g., by a signal generated from a label attached to the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 nucleic acid, polypeptide, or antibody.

The capture probes can be a set of nucleic acids from a selected sample, e.g., a sample of nucleic acids derived from a control or non-stimulated tissue or cell.

The method can include contacting the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 nucleic acid, polypeptide, or antibody with a first array having a plurality of capture probes and a second array having a different plurality of capture probes. The results of each hybridization can be compared, e.g., to analyze differences in expression between a first and second sample. The first plurality of capture probes can be from a control sample, e.g., a wild type, normal, or non-diseased, non-stimulated, sample, e.g., a biological fluid, tissue, or cell sample. The second plurality of capture probes can be from an experimental sample, e.g., a mutant type, at risk, disease-state or disorder-state, or stimulated, sample, e.g., a biological fluid, tissue, or cell sample.

The plurality of capture probes can be a plurality of nucleic acid probes each of which specifically hybridizes, with an allele of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843. Such methods can be used to diagnose a subject, e.g., to evaluate risk for a disease or disorder, to evaluate suitability of a selected treatment for a subject, to evaluate whether a subject has a disease or disorder.

The method can be used to detect SNPs, as described above.

In another aspect, the invention features, a method of analyzing 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843, e.g., analyzing structure, function, or relatedness to other nucleic acid or amino acid sequences. The method includes: providing a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 nucleic acid or amino acid sequence; comparing the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 sequence with one or more preferably a plurality of sequences from a collection of sequences, e.g., a nucleic acid or protein sequence database; to thereby analyze 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843.

The method can include evaluating the sequence identity between a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 sequence and a database sequence. The method can be performed by accessing the database at a second site, e.g., over the internet. Preferred databases include GenBank™ and SwissProt.

In another aspect, the invention features, a set of oligonucleotides, useful, e.g., for identifying SNP's, or identifying specific alleles of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843. The set includes a plurality of oligonucleotides, each of which has a different nucleotide at an interrogation position, e.g., an SNP or the site of a mutation. In a preferred embodiment, the oligonucleotides of the plurality identical in sequence with one another (except for differences in length). The oligonucleotides can be provided with differential labels, such that an oligonucleotide which hybridizes to one allele provides a signal that is distinguishable from an oligonucleotides which hybridizes to a second allele.

The sequences of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 molecules are provided in a variety of mediums to facilitate use thereof. A sequence can be provided as a manufacture, other than an isolated nucleic acid or amino acid molecule, which contains a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 molecule. Such a manufacture can provide a nucleotide or amino acid sequence, e.g., an open reading frame, in a form which allows examination of the manufacture using means not directly applicable to examining the nucleotide or amino acid sequences, or a subset thereof, as they exist in nature or in purified form.

A 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 nucleotide or amino acid sequence can be recorded on computer readable media. As used herein, “computer readable media” refers to any medium that can be read and accessed directly by a computer. Such media include, but are not limited to: magnetic storage media, such as floppy discs, hard disc storage medium, and magnetic tape; optical storage media such as compact disc and CD-ROM; electrical storage media such as RAM, ROM, EPROM, EEPROM, and the like; and general hard disks and hybrids of these categories such as magnetic/optical storage media. The medium is adapted or configured for having thereon 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 sequence information of the present invention.

As used herein, the term “electronic apparatus” is intended to include any suitable computing or processing apparatus of other device configured or adapted for storing data or information. Examples of electronic apparatus suitable for use with the present invention include stand-alone computing apparatus; networks, including a local area network (LAN), a wide area network (WAN) Internet, Intranet, and Extranet; electronic appliances such as personal digital assistants (PDAs), cellular phones, pagers, and the like; and local and distributed processing systems.

As used herein, “recorded” refers to a process for storing or encoding information on the electronic apparatus readable medium. Those skilled in the art can readily adopt any of the presently known methods for recording information on known media to generate manufactures comprising the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 sequence information.

A variety of data storage structures are available to a skilled artisan for creating a computer readable medium having recorded thereon a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 nucleotide or amino acid sequence of the present invention. The choice of the data storage structure will generally be based on the means chosen to access the stored information. In addition, a variety of data processor programs and formats can be used to store the nucleotide sequence information of the present invention on computer readable medium. The sequence information can be represented in a word processing text file, formatted in commercially-available software such as WordPerfect and Microsoft Word, or represented in the form of an ASCII file, stored in a database application, such as DB2, Sybase, Oracle, or the like. The skilled artisan can readily adapt any number of data processor structuring formats (e.g., text file or database) in order to obtain computer readable medium having recorded thereon the nucleotide sequence information of the present invention.

By providing the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 nucleotide or amino acid sequences of the invention in computer readable form, the skilled artisan can routinely access the sequence information for a variety of purposes. For example, one skilled in the art can use the nucleotide or amino acid sequences of the invention in computer readable form to compare a target sequence or target structural motif with the sequence information stored within the data storage means. A search is used to identify fragments or regions of the sequences of the invention which match a particular target sequence or target motif.

The present invention therefore provides a medium for holding instructions for performing a method for determining whether a subject has a arginine methyltransferase, glycosyltransferase, gamra-glutamyltraspeptidase, phosphoribosylglycinamide transferase, acyltransferase, acyl-CoA dehydrogenase, fatty acid amide hydrolase, aminotransferase, zinc carboxypeptidase, protein kinase, DEAD helicase, short-chain dehydrogenase/reductase or phosphatase-associated or another 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843-associated disease or disorder or a pre-disposition to a arginine methyltransferase, glycosyltransferase, gamma-glutamyltraspeptidase, phosphoribosylglycinamide transferase, acyltransferase, acyl-CoA dehydrogenase, fatty acid amide hydrolase, aminotransferase, zinc carboxypeptidase, protein kinase, DEAD helicase, short-chain dehydrogenase/reductase or phosphatase-associated or another 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843-associated disease or disorder, wherein the method comprises the steps of determining 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 sequence information associated with the subject and based on the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 sequence information, determining whether the subject has a arginine methyltransferase, glycosyltransferase, gamma-glutamyltraspeptidase, phosphoribosylglycinamide transferase, acyltransferase, acyl-CoA dehydrogenase, fatty acid amide hydrolase, aminotransferase, zinc carboxypeptidase, protein kinase, DEAD helicase, short-chain dehydrogenase/reductase or phosphatase-associated or another 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843-associated disease or disorder and/or recommending a particular treatment for the disease, disorder, or pre-disease condition.

The present invention further provides in an electronic system and/or in a network, a method for determining whether a subject has a arginine methyltransferase, glycosyltransferase, gamma-glutamyltraspeptidase, phosphoribosylglycinamide transferase, acyltransferase, acyl-CoA dehydrogenase, fatty acid amide hydrolase, aminotransferase, zinc carboxypeptidase, protein kinase, DEAD helicase, short-chain dehydrogenase/reductase or phosphatase-associated or another 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843-associated disease or disorder or a pre-disposition to a disease associated with 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843, wherein the method comprises the steps of determining 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 sequence information associated with the subject, and based on the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 sequence information, determining whether the subject has a arginine methyltransferase, glycosyltransferase, gamma-glutamyltraspeptidase, phosphoribosylglycinamide transferase, acyltransferase, acyl-CoA dehydrogenase, fatty acid amide hydrolase, aminotransferase, zinc carboxypeptidase, protein kinase, DEAD helicase, short-chain dehydrogenase/reductase or phosphatase-associated or another 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843-associated disease or disorder or a pre-disposition to a arginine methyltransferase, glycosyltransferase, gamma-glutamyltraspeptidase, phosphoribosylglycinamide transferase, acyltransferase, acyl-CoA dehydrogenase, fatty acid amide hydrolase, aminotransferase, zinc carboxypeptidase, protein kinase, DEAD helicase, short-chain dehydrogenase/reductase or phosphatase-associated or another 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843-associated disease or disorder, and/or recommending a particular treatment for the disease, disorder, or pre-disease condition. The method may further comprise the step of receiving phenotypic information associated with the subject and/or acquiring from a network phenotypic information associated with the subject.

The present invention also provides in a network, a method for determining whether a subject has a arginine methyltransferase, glycosyltransferase, gamma-glutamyltraspeptidase, phosphoribosylglycinamide transferase, acyltransferase, acyl-CoA dehydrogenase, fatty acid amide hydrolase, aminotransferase, zinc carboxypeptidase, protein kinase, DEAD helicase, short-chain dehydrogenase/reductase or phosphatase-associated or another 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843-associated disease or disorder or a pre-disposition to a arginine methyltransferase, glycosyltransferase, gamma-glutamyltraspeptidase, phosphoribosylglycinamide transferase, acyltransferase, acyl-CoA dehydrogenase, fatty acid amide hydrolase, aminotransferase, zinc carboxypeptidase, protein kinase, DEAD helicase, short-chain dehydrogenase/reductase or phosphatase-associated or another 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843-associated disease or disorder, said method comprising the steps of receiving 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 sequence information from the subject and/or information related thereto, receiving phenotypic information associated with the subject, acquiring information from the network corresponding to 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 and/or corresponding to a arginine methyltransferase, glycosyltransferase, gamma-glutamyltraspeptidase, phosphoribosylglycinamide transferase, acyltransferase, acyl-CoA dehydrogenase, fatty acid amide hydrolase, aminotransferase, zinc carboxypeptidase, protein kinase, DEAD helicase, short-chain dehydrogenase/reductase or phosphatase-associated or another 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843-associated disease or disorder, and based on one or more of the phenotypic information, the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 information (e.g., sequence information and/or information related thereto), and the acquired information, determining whether the subject has a arginine methyltransferase, glycosyltransferase, gamma-glutamyltraspeptidase, phosphoribosylglycinamide transferase, acyltransferase, acyl-CoA dehydrogenase, fatty acid amide hydrolase, aminotransferase, zinc carboxypeptidase, protein kinase, DEAD helicase, short-chain dehydrogenase/reductase or phosphatase-associated or another 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843-associated disease or disorder or a pre-disposition to a arginine methyltransferase, glycosyltransferase, gamma-glutamyltraspeptidase, phosphoribosylglycinamide transferase, acyltransferase, acyl-CoA dehydrogenase, fatty acid amide hydrolase, aminotransferase, zinc carboxypeptidase, protein kinase, DEAD helicase, short-chain dehydrogenase/reductase or phosphatase-associated or another 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843-associated disease or disorder. The method may further comprise the step of recommending a particular treatment for the disease, disorder, or pre-disease condition.

The present invention also provides a business method for determining whether a subject has a arginine methyltransferase, glycosyltransferase, gamma-glutamyltraspeptidase, phosphoribosylglycinamide transferase, acyltransferase, acyl-CoA dehydrogenase, fatty acid amide hydrolase, aminotransferase, zinc carboxypeptidase, protein kinase, DEAD helicase, short-chain dehydrogenase/reductase or phosphatase-associated or another 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 -associated disease or disorder or a pre-disposition to a arginine methyltransferase, glycosyltransferase, gamma-glutamyltraspeptidase, phosphoribosylglycinamide transferase, acyltransferase, acyl-CoA dehydrogenase, fatty acid amide hydrolase, aminotransferase, zinc carboxypeptidase, protein kinase, DEAD helicase, short-chain dehydrogenase/reductase or phosphatase-associated or another 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843-associated disease or disorder, said method comprising the steps of receiving information related to 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 (e.g., sequence information and/or information related thereto), receiving phenotypic information associated with the subject, acquiring information from the network related to 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 and/or related to a arginine methyltransferase, glycosyltransferase, gamma-glutamyltraspeptidase, phosphoribosylglycinamide transferase, acyltransferase, acyl-CoA dehydrogenase, fatty acid amide hydrolase, aminotransferase, zinc carboxypeptidase, protein kinase, DEAD helicase, short-chain dehydrogenase/reductase or phosphatase-associated or another 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843-associated disease or disorder, and based on one or more of the phenotypic information, the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 information, and the acquired information, determining whether the subject has a arginine methyltransferase, glycosyltransferase, gamma-glutamyltraspeptidase, phosphoribosylglycinamide transferase, acyltransferase, acyl-CoA dehydrogenase, fatty acid amide hydrolase, aminotransferase, zinc carboxypeptidase, protein kinase, DEAD helicase, short-chain dehydrogenase/reductase or phosphatase-associated or another 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843-associated disease or disorder or a pre-disposition to a arginine methyltransferase, glycosyltransferase, gamma-glutamyltraspeptidase, phosphoribosylglycinamide transferase, acyltransferase, acyl-CoA dehydrogenase, fatty acid amide hydrolase, aminotransferase, zinc carboxypeptidase, protein kinase, DEAD helicase, short-chain dehydrogenase/reductase or phosphatase-associated or another 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843-associated disease or disorder. The method may further comprise the step of recommending a particular treatment for the disease, disorder, or pre-disease condition.

The invention also includes an array comprising a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 sequence of the present invention. The array can be used to assay expression of one or more genes in the array. In one embodiment, the array can be used to assay gene expression in a tissue to ascertain tissue specificity of genes in the array. In this manner, up to about 7600 genes can be simultaneously assayed for expression, one of which can be 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843. This allows a profile to be developed showing a battery of genes specifically expressed in one or more tissues.

In addition to such qualitative information, the invention allows the quantitation of gene expression. Thus, not only tissue specificity, but also the level of expression of a battery of genes in the tissue if ascertainable. Thus, genes can be grouped on the basis of their tissue expression per se and level of expression in that tissue. This is useful, for example, in ascertaining the relationship of gene expression in that tissue. Thus, one tissue can be perturbed and the effect on gene expression in a second tissue can be determined. In this context, the effect of one cell type on another cell type in response to a biological stimulus can be determined. In this context, the effect of one cell type on another cell type in response to a biological stimulus can be determined. Such a determination is useful, for example, to know the effect of cell-cell interaction at the level of gene expression. If an agent is administered therapeutically to treat one cell type but has an undesirable effect on another cell type, the invention provides an assay to determine the molecular basis of the undesirable effect and thus provides the opportunity to co-administer a counteracting agent or otherwise treat the undesired effect. Similarly, even within a single cell type, undesirable biological effects can be determined at the molecular level. Thus, the effects of an agent on expression of other than the target gene can be ascertained and counteracted.

In another embodiment, the array can be used to monitor the time course of expression of one or more genes in the array. This can occur in various biological contexts, as disclosed herein, for example development of an arginine methyltransferase, glycosyltransferase, gamma-glutamyltraspeptidase, phosphoribosylglycinamide transferase, acyltransferase, acyl-CoA dehydrogenase, fatty acid amide hydrolase, aminotransferase, zinc carboxypeptidase, protein kinase, DEAD helicase, short-chain dehydrogenase/reductase or phosphatase-associated or another 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843-associated disease or disorder, progression of an arginine methyltransferase, glycosyltransferase, gamma-glutamyltraspeptidase, phosphoribosylglycinamide transferase, acyltransferase, acyl-CoA dehydrogenase, fatty acid amide hydrolase, aminotransferase, zinc carboxypeptidase, protein kinase, DEAD helicase, short-chain dehydrogenase/reductase or phosphatase-associated or another 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843-associated disease or disorder, and processes, such a cellular transformation associated with the arginine methyltransferase, glycosyltransferase, gamma-glutamyltraspeptidase, phosphoribosylglycinamide transferase, acyltransferase, acyl-CoA dehydrogenase, fatty acid amide hydrolase, aminotransferase, zinc carboxypeptidase, protein kinase, DEAD helicase, short-chain dehydrogenase/reductase or phosphatase-associated or another 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843-associated disease or disorder.

The array is also useful for ascertaining the effect of the expression of a gene on the expression of other genes in the same cell or in different cells (e.g., acertaining the effect of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 expression on the expression of other genes). This provides, for example, for a selection of alternate molecular targets for therapeutic intervention if the ultimate or downstream target cannot be regulated.

The array is also useful for ascertaining differential expression patterns of one or more genes in normal and abnormal cells. This provides a battery of genes (e.g., including 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843) that could serve as a molecular target for diagnosis or therapeutic intervention.

As used herein, a “target sequence” can be any DNA or amino acid sequence of six or more nucleotides or two or more amino acids. A skilled artisan can readily recognize that the longer a target sequence is, the less likely a target sequence will be present as a random occurrence in the database. Typical sequence lengths of a target sequence are from about 10 to 100 amino acids or from about 30 to 300 nucleotide residues. However, it is well recognized that commercially important fragments, such as sequence fragments involved in gene expression and protein processing, may be of shorter length.

Computer software is publicly available which allows a skilled artisan to access sequence information provided in a computer readable medium for analysis and comparison to other sequences. A variety of known algorithms are disclosed publicly and a variety of commercially available software for conducting search means are and can be used in the computer-based systems of the present invention. Examples of such software include, but are not limited to, MacPattern (EMBL), BLASTN and BLASTX (NCBI).

Thus, the invention features a method of making a computer readable record of a sequence of a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 sequence which includes recording the sequence on a computer readable matrix. In a preferred embodiment the record includes one or more of the following: identification of an ORF; identification of a domain, region, or site; identification of the start of transcription; identification of the transcription terminator; the full length amino acid sequence of the protein, or a mature form thereof; the 5′ end of the translated region.

In another aspect, the invention features a method of analyzing a sequence. The method includes: providing a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 sequence, or record, in computer readable form; comparing a second sequence to the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 sequence; thereby analyzing a sequence. Comparison can include comparing to sequences for sequence identity or determining if one sequence is included within the other, e.g., determining if the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 sequence includes a sequence being compared. In a preferred embodiment the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 or second sequence is stored on a first computer, e.g., at a first site and the comparison is performed, read, or recorded on a second computer, e.g., at a second site. E.g., the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 or second sequence can be stored in a public or proprietary database in one computer, and the results of the comparison performed, read, or recorded on a second computer. In a preferred embodiment the record includes one or more of the following: identification of an ORF; identification of a domain, region, or site; identification of the start of transcription; identification of the transcription terminator; the full length amino acid sequence of the protein, or a mature form thereof; the 5′ end of the translated region.

EXEMPLIFICATION

Example 1

Tissue Distribution of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 mRNA [0862]
Northern blot hybridizations with various RNA samples can be performed under standard conditions and washed under stringent conditions, i.e., 0.2×SSC at 65° C. A DNA probe corresponding to all or a portion of the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 cDNA (SEQ ID NO:1, 3, 4, 6, 7, 9, 10, 12, 13, 15, 16, 18, 53, 55, 61, 63, 67, 69, 78, 80, 88, 90, 100, 102, 113, 115, 122, 124, 129 or 131) or 26199, 33530, 339499 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 cDNA can be used. The DNA was radioactively labeled with [0863] ³²P-dCTP using the Prime-It Kit (Stratagene, La Jolla, Calif.) according to the instructions of the supplier. Filters containing mRNA from mouse hematopoietic and endocrine tissues, and cancer cell lines (Clontech, Palo Alto, Calif.) can be probed in ExpressHyb hybridization solution (Clontech) and washed at high stringency according to manufacturer's recommendations.

Example 2

Recombinant Expression of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 in Bacterial Cells [0864]
In this example, 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 is expressed as a recombinant glutathione-S-transferase (GST) fusion polypeptide in [0865] E. coli and the fusion polypeptide is isolated and characterized. Specifically, 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 is fused to GST and this fusion polypeptide is expressed in E. coli, e.g., strain PEB 199. Expression of the GST-26199, -33530, -33949, -47148, -50226, -58764, -62113, -32144, -32235, -23565, -13305, -14911, -86216, -25206 or -8843 fusion protein in PEB199 is induced with IPTG. The recombinant fusion polypeptide is purified from crude bacterial lysates of the induced PEB 199 strain by affinity chromatography on glutathione beads. Using polyacrylamide gel electrophoretic analysis of the polypeptide purified from the bacterial lysates, the molecular weight of the resultant fusion polypeptide is determined.

Example 3

Expression of Recombinant 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 Protein in COS Cells [0866]
To express the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 gene in COS cells, the pcDNA/Amp vector by Invitrogen Corporation (San Diego, Calif.) is used. This vector contains an SV40 origin of replication, an ampicillin resistance gene, an [0867] E. coli replication origin, a CMV promoter followed by a polylinker region, and an SV40 intron and polyadenylation site. A DNA fragment encoding the entire 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 protein and an HA tag (Wilson et al. (1984) Cell 37:767) or a FLAG tag fused in-frame to its 3′ end of the fragment is cloned into the polylinker region of the vector, thereby placing the expression of the recombinant protein under the control of the CMV promoter.
To construct the plasmid, the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 DNA sequence is amplified by PCR using two primers. The 5′ primer contains the restriction site of interest followed by approximately twenty nucleotides of the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 coding sequence starting from the initiation codon; the 3′ end sequence contains complementary sequences to the other restriction site of interest, a translation stop codon, the HA tag or FLAG tag and the last 20 nucleotides of the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 coding sequence. The PCR amplified fragment and the pCDNA/Amp vector are digested with the appropriate restriction enzymes and the vector is dephosphorylated using the CIAP enzyme (New England Biolabs, Beverly, Mass.). Preferably the two restriction sites chosen are different so that the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 gene is inserted in the correct orientation. The ligation mixture is transformed into [0868] E. coli cells (strains HB101, DH5α, SURE, available from Stratagene Cloning Systems, La Jolla, Calif., can be used), the transformed culture is plated on ampicillin media plates, and resistant colonies are selected. Plasmid DNA is isolated from transformants and examined by restriction analysis for the presence of the correct fragment.
COS cells are subsequently transfected with the 26199-, 33530-, 33949-, 47148-, 50226-, 58764-, 62113-, 32144-, 32235-, 23565-, 13305-, 14911-, 86216-, 25206- or 8843-pcDNA/Amp plasmid DNA using the calcium phosphate or calcium chloride co-precipitation methods, DEAE-dextran-mediated transfection, lipofection, or electroporation. Other suitable methods for transfecting host cells can be found in Sambrook, J., Fritsh, E. F., and Maniatis, T. [0869] Molecular Cloning: A Laboratory Manual. 2nd, ed., Cold Spring Harbor Laboratory, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1989. The expression of the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 polypeptide is detected by radiolabelling (35S-methionine or ³⁵S-cysteine available from NEN, Boston, Mass., can be used) and immunoprecipitation (Harlow, E. and Lane, D. Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1988) using an HA specific monoclonal antibody. Briefly, the cells are labeled for 8 hours with ³⁵S-methionine (or ³⁵S-cysteine). The culture media are then collected and the cells are lysed using detergents (RIPA buffer, 150 mM NaCl, 1% NP-40, 0.1% SDS, 0.5% DOC, 50 mM Tris, pH 7.5). Both the cell lysate and the culture media are precipitated with an HA specific monoclonal antibody. Precipitated polypeptides are then analyzed by SDS-PAGE.
Alternatively, DNA containing the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 coding sequence is cloned directly into the polylinker of the pCDNA/Amp vector using the appropriate restriction sites. The resulting plasmid is transfected into COS cells in the manner described above, and the expression of the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 polypeptide is detected by radiolabelling and immunoprecipitation using a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 specific monoclonal antibody. [0870]

Exmaple 4

TaqMan Analysis of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 [0871]
Human 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 expression was measured by TaqMan® quantitative PCR (Perkin Elmer Applied Biosystems) in cDNA prepared from a variety of normal and diseased (e.g., cancerous) human tissues or cell lines. [0872]
Probes were designed by PrimerExpress software (PE Biosystems) based on the sequence of the human 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 gene. Each human 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 gene probe was labeled using FAM (6-carboxyfluorescein), and the β2-microglobulin reference probe was labeled with a different fluorescent dye, VIC. The differential labeling of the target gene and internal reference gene thus enabled measurement in same well. Forward and reverse primers and the probes for both β2-microglobulin and target gene were added to the TaqMan® Universal PCR Master Mix (PE Applied Biosystems). Although the final concentration of primer and probe could vary, each was internally consistent within a given experiment. A typical experiment contained 200 nM of forward and reverse primers plus 100 nM probe for β-2 microglobulin and 600 nM forward and reverse primers plus 200 nM probe for the target gene. TaqMan matrix experiments were carried out on an ABI PRISM 7700 Sequence Detection System (PE Applied Biosystems). The thermal cycler conditions were as follows: hold for 2 min at 50° C. and 10 min at 95° C., followed by two-step PCR for 40 cycles of 95° C. for 15 sec followed by 60° C. for 1 min. [0873]
The following method was used to quantitatively calculate human 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 gene expression in the various tissues relative to β-2 microglobulin expression in the same tissue. The threshold cycle (Ct) value is defined as the cycle at which a statistically significant increase in fluorescence is detected. A lower Ct value is indicative of a higher mRNA concentration. The Ct value of the human 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 gene is normalized by subtracting the Ct value of the β-2 microglobulin gene to obtain a [0874] _ΔCt value using the following formula: _ΔCt=Ct_sample−Ct_{β-2 microglobulin}. Expression is then calibrated against a cDNA sample showing a comparatively low level of expression of the human 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 gene. The _ΔCt value for the calibrator sample is then subtracted from _ΔCt for each tissue sample according to the following formula: _ΔΔCt=_ΔCt-_sample−_ΔCt-_calibrator. Relative expression is then calculated using the arithmetic formula given by 2^−ΔΔCt.

Example 5

In Situ Hybridization of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 [0875]
The following describes the tissue distribution of 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 mRNA, as may be determined by in situ hybridization analysis using oligonucleotide probes based on the human G2RF sequence. [0876]
For in situ analysis, various tissues, e.g. tissues obtained from brain, are first frozen on dry ice. Ten-micrometer-thick sections of the tissues are postfixed with 4% formaldehyde in DEPC treated 1× phosphate-buffered saline at room temperature for 10 minutes before being rinsed twice in DEPC 1× phosphate-buffered saline and once in 0.1 M triethanolamine-HCl (pH 8.0). Following incubation in 0.25% acetic anhydride-0.1 M triethanolamine-HCl for 10 minutes, sections are rinsed in DEPC 2×SSC (1×SSC is 0.15M NaCl plus 0.015M sodium citrate). Tissue is then dehydrated through a series of ethanol washes, incubated in 100% chloroform for 5 minutes, and then rinsed in 100% ethanol for 1 minute and 95% ethanol for 1 minute and allowed to air dry. [0877]
Hybridizations are performed with [0878] ³⁵S-radiolabeled (5×10⁷cpm/ml) cRNA probes. Probes are incubated in the presence of a solution containing 600 mM NaCl, 10 mM Tris (pH 7.5), 1 mM EDTA, 0.01% sheared salmon sperm DNA, 0.01% yeast tRNA, 0.05% yeast total RNA type X1, 1× Denhardt's solution, 50% formamide, 10% dextran sulfate, 100 mM dithiothreitol, 0.1% sodium dodecyl sulfate (SDS), and 0.1% sodium thiosulfate for 18 hours at 55° C.
After hybridization, slides are washed with 2×SSC. Sections are then sequentially incubated at 37° C. in TNE (a solution containing 10 mM Tris-HCl (pH 7.6), 500 mM NaCl, and 1 mM EDTA), for 10 minutes, in TNE with 10 μg of RNase A per ml for 30 minutes, and finally in TNF for 10 minutes. Slides are then rinsed with 2×SSC at room temperature, washed with 2×SSC at 50° C. for 1 hour, washed with 0.2×SSC at 55° C. for 1 hour, and 0.2×SSC at 60° C. for 1 hour. Sections are then dehydrated rapidly through serial ethanol-0.3 M sodium acetate concentrations before being air dried and exposed to Kodak Biomax MR scientific imaging film for 24 hours and subsequently dipped in NB-2 photoemulsion and exposed at 4° C. for 7 days before being developed and counter stained. [0879]
The contents of all references, patents and published patent applications cited throughout this application are incorporated herein by reference. [0880]
Equivalents [0881]
Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. [0882]
1 136 1 1828 DNA Homo sapiens CDS (56)...(745) 1 gcggtgcgcg ttgatgtgac gtccctgcgc gcgccgcttt ctgttgccgg gcgca atg 58 Met 1 gcg gat acg ctg gag tcc tcg ctg gag gac cca ctg cgg agc ttt gtg 106 Ala Asp Thr Leu Glu Ser Ser Leu Glu Asp Pro Leu Arg Ser Phe Val 5 10 15 cga gtt ttg gag aag cgg gat ggt aca gtg cta cga cta cag cag tat 154 Arg Val Leu Glu Lys Arg Asp Gly Thr Val Leu Arg Leu Gln Gln Tyr 20 25 30 agc tcc ggt ggc gtg ggt tgc gtt gtg tgg gac gct gcc att gtc ctt 202 Ser Ser Gly Gly Val Gly Cys Val Val Trp Asp Ala Ala Ile Val Leu 35 40 45 tct aaa tac ctg gaa acg ccc gag ttt tct ggc gac ggg gcc cac gcg 250 Ser Lys Tyr Leu Glu Thr Pro Glu Phe Ser Gly Asp Gly Ala His Ala 50 55 60 65 ctg agc cgg cgg tcg gtg ctg gag ctg ggt tcg ggc acc ggg gcc gtg 298 Leu Ser Arg Arg Ser Val Leu Glu Leu Gly Ser Gly Thr Gly Ala Val 70 75 80 ggg ctc atg gct gct acc ctc ggg gct gat gtt gta gtc acc gat ctt 346 Gly Leu Met Ala Ala Thr Leu Gly Ala Asp Val Val Val Thr Asp Leu 85 90 95 gag gaa ttg caa gac ttg ctg aag atg aat att aat atg aac aag cat 394 Glu Glu Leu Gln Asp Leu Leu Lys Met Asn Ile Asn Met Asn Lys His 100 105 110 ctt gtc act ggt tct gtt caa gcc aag gta ctg aaa tgg ggg gaa gaa 442 Leu Val Thr Gly Ser Val Gln Ala Lys Val Leu Lys Trp Gly Glu Glu 115 120 125 ata gaa ggc ttt cct tct cca ccc gac ttc ata ctg atg gcc gac tgc 490 Ile Glu Gly Phe Pro Ser Pro Pro Asp Phe Ile Leu Met Ala Asp Cys 130 135 140 145 ata tac tat gaa gag tct ttg gag cca ttg ctg aaa act cta aaa gat 538 Ile Tyr Tyr Glu Glu Ser Leu Glu Pro Leu Leu Lys Thr Leu Lys Asp 150 155 160 atc agc gga ttt gaa act tgt att ata tgt tgt tat gaa caa cga aca 586 Ile Ser Gly Phe Glu Thr Cys Ile Ile Cys Cys Tyr Glu Gln Arg Thr 165 170 175 atg ggg aaa aat cca gaa att gag aaa aaa tat ttt gag ctc ctt cag 634 Met Gly Lys Asn Pro Glu Ile Glu Lys Lys Tyr Phe Glu Leu Leu Gln 180 185 190 cta gat ttt gac ttt gaa aaa att cct ttg gaa aaa cat gat gaa gag 682 Leu Asp Phe Asp Phe Glu Lys Ile Pro Leu Glu Lys His Asp Glu Glu 195 200 205 tat cga agt gaa gat att cat att ata tac atc aga aag aaa aaa tcg 730 Tyr Arg Ser Glu Asp Ile His Ile Ile Tyr Ile Arg Lys Lys Lys Ser 210 215 220 225 aaa ttt cca tcg tga agcctttaat catcttaccc aaggctctaa caacctgggt 785 Lys Phe Pro Ser * aaactaaaga tgtgaataga gcatgtgaat acagcatggg aagattgtgt tcacagattt 845 ttttttccgg cacgtcctta gagatccgat gtatagatga tgaccaccag gggctgtctg 905 caatacgaaa aattcctgct tgcctgcctg cctgccaatg gccctgaatc cagcttaggt 965 tacttagttc agcatcaagt tcttcttaaa tgttgggaat cagttattca aataaaaatt 1025 ggtattgagg cgaggctgaa atctgccaaa aacagccaac atgatttgac tggacctttt 1085 acaggaagct agagataaat ttctgaagag aactatctgc tattatataa taatgtttta 1145 aattcaaact agtaaatttt agtttgtctt cagagtttaa aaggttttca ttttgtacat 1205 aattatacaa tatttatcat ttgtattttc cacttcattt cttttaaaat atctttaata 1265 ctgaaatgtt cttttaattt taaaaagaac tgagatattg tcttgtatac ttatattggc 1325 caaagttttc ttttcctcca ccatacatat ctatgtgatt taatcagtaa attgtactgt 1385 aatgagttgc taagaagaca aatcagaaga ttggaggaac aaaagatatc tttacagatt 1445 tttcatttga gcatggagtg agaatagaaa gaccagtttc aggcttatgc acttacgtgg 1505 ctcatgcact ttatgtatat gttccaggaa aatctggctt aaaaatactg gtattgttta 1565 catgaagcag tgaaaggttt ttgaataact acaaatgtag ttctatatgt atataccaaa 1625 tgaatttctg ttctgtgtct ctctgtttta tgttatgaag ccattcgctc atatacaata 1685 atctgtcaag gactttaatc atacttgttc caaagagtag taaccggatg gaattctggt 1745 atttacaggc attggtgcta gatggtacat tttatgtgtt aaaataaaca ttgtttttga 1805 gaaaaaaaaa aaaaaaaars gaa 1828 2 229 PRT Homo sapiens 2 Met Ala Asp Thr Leu Glu Ser Ser Leu Glu Asp Pro Leu Arg Ser Phe 1 5 10 15 Val Arg Val Leu Glu Lys Arg Asp Gly Thr Val Leu Arg Leu Gln Gln 20 25 30 Tyr Ser Ser Gly Gly Val Gly Cys Val Val Trp Asp Ala Ala Ile Val 35 40 45 Leu Ser Lys Tyr Leu Glu Thr Pro Glu Phe Ser Gly Asp Gly Ala His 50 55 60 Ala Leu Ser Arg Arg Ser Val Leu Glu Leu Gly Ser Gly Thr Gly Ala 65 70 75 80 Val Gly Leu Met Ala Ala Thr Leu Gly Ala Asp Val Val Val Thr Asp 85 90 95 Leu Glu Glu Leu Gln Asp Leu Leu Lys Met Asn Ile Asn Met Asn Lys 100 105 110 His Leu Val Thr Gly Ser Val Gln Ala Lys Val Leu Lys Trp Gly Glu 115 120 125 Glu Ile Glu Gly Phe Pro Ser Pro Pro Asp Phe Ile Leu Met Ala Asp 130 135 140 Cys Ile Tyr Tyr Glu Glu Ser Leu Glu Pro Leu Leu Lys Thr Leu Lys 145 150 155 160 Asp Ile Ser Gly Phe Glu Thr Cys Ile Ile Cys Cys Tyr Glu Gln Arg 165 170 175 Thr Met Gly Lys Asn Pro Glu Ile Glu Lys Lys Tyr Phe Glu Leu Leu 180 185 190 Gln Leu Asp Phe Asp Phe Glu Lys Ile Pro Leu Glu Lys His Asp Glu 195 200 205 Glu Tyr Arg Ser Glu Asp Ile His Ile Ile Tyr Ile Arg Lys Lys Lys 210 215 220 Ser Lys Phe Pro Ser 225 3 690 DNA Homo sapiens CDS (1)...(690) 3 atg gcg gat acg ctg gag tcc tcg ctg gag gac cca ctg cgg agc ttt 48 Met Ala Asp Thr Leu Glu Ser Ser Leu Glu Asp Pro Leu Arg Ser Phe 1 5 10 15 gtg cga gtt ttg gag aag cgg gat ggt aca gtg cta cga cta cag cag 96 Val Arg Val Leu Glu Lys Arg Asp Gly Thr Val Leu Arg Leu Gln Gln 20 25 30 tat agc tcc ggt ggc gtg ggt tgc gtt gtg tgg gac gct gcc att gtc 144 Tyr Ser Ser Gly Gly Val Gly Cys Val Val Trp Asp Ala Ala Ile Val 35 40 45 ctt tct aaa tac ctg gaa acg ccc gag ttt tct ggc gac ggg gcc cac 192 Leu Ser Lys Tyr Leu Glu Thr Pro Glu Phe Ser Gly Asp Gly Ala His 50 55 60 gcg ctg agc cgg cgg tcg gtg ctg gag ctg ggt tcg ggc acc ggg gcc 240 Ala Leu Ser Arg Arg Ser Val Leu Glu Leu Gly Ser Gly Thr Gly Ala 65 70 75 80 gtg ggg ctc atg gct gct acc ctc ggg gct gat gtt gta gtc acc gat 288 Val Gly Leu Met Ala Ala Thr Leu Gly Ala Asp Val Val Val Thr Asp 85 90 95 ctt gag gaa ttg caa gac ttg ctg aag atg aat att aat atg aac aag 336 Leu Glu Glu Leu Gln Asp Leu Leu Lys Met Asn Ile Asn Met Asn Lys 100 105 110 cat ctt gtc act ggt tct gtt caa gcc aag gta ctg aaa tgg ggg gaa 384 His Leu Val Thr Gly Ser Val Gln Ala Lys Val Leu Lys Trp Gly Glu 115 120 125 gaa ata gaa ggc ttt cct tct cca ccc gac ttc ata ctg atg gcc gac 432 Glu Ile Glu Gly Phe Pro Ser Pro Pro Asp Phe Ile Leu Met Ala Asp 130 135 140 tgc ata tac tat gaa gag tct ttg gag cca ttg ctg aaa act cta aaa 480 Cys Ile Tyr Tyr Glu Glu Ser Leu Glu Pro Leu Leu Lys Thr Leu Lys 145 150 155 160 gat atc agc gga ttt gaa act tgt att ata tgt tgt tat gaa caa cga 528 Asp Ile Ser Gly Phe Glu Thr Cys Ile Ile Cys Cys Tyr Glu Gln Arg 165 170 175 aca atg ggg aaa aat cca gaa att gag aaa aaa tat ttt gag ctc ctt 576 Thr Met Gly Lys Asn Pro Glu Ile Glu Lys Lys Tyr Phe Glu Leu Leu 180 185 190 cag cta gat ttt gac ttt gaa aaa att cct ttg gaa aaa cat gat gaa 624 Gln Leu Asp Phe Asp Phe Glu Lys Ile Pro Leu Glu Lys His Asp Glu 195 200 205 gag tat cga agt gaa gat att cat att ata tac atc aga aag aaa aaa 672 Glu Tyr Arg Ser Glu Asp Ile His Ile Ile Tyr Ile Arg Lys Lys Lys 210 215 220 tcg aaa ttt cca tcg tga 690 Ser Lys Phe Pro Ser * 225 4 1408 DNA Homo sapiens CDS (36)...(1286) 4 ccagtgtggt ggaattcagt tgcggctcca gggcc atg gcg gag gag cag gcc 53 Met Ala Glu Glu Gln Ala 1 5 cgg gaa cgg gac tcg gtt ccc aag ccg tcg gtg ctg ttc ctc cac cca 101 Arg Glu Arg Asp Ser Val Pro Lys Pro Ser Val Leu Phe Leu His Pro 10 15 20 gac ctg ggc gtg ggc ggc gct gag cgg ctg gtg ttg gac gcg gcg ctg 149 Asp Leu Gly Val Gly Gly Ala Glu Arg Leu Val Leu Asp Ala Ala Leu 25 30 35 gcg ctg cag gcg cgc ggg tgt agc gtg aag atc tgg aca gcg cac tac 197 Ala Leu Gln Ala Arg Gly Cys Ser Val Lys Ile Trp Thr Ala His Tyr 40 45 50 gac ccg ggc cac tgt ttc gcc gag agc cgc gag cta ccg gtg cgc tgt 245 Asp Pro Gly His Cys Phe Ala Glu Ser Arg Glu Leu Pro Val Arg Cys 55 60 65 70 gcc ggg gac tgg ctg ccg cga ggc ctg ggc tgg ggc ggc cgc ggc gcc 293 Ala Gly Asp Trp Leu Pro Arg Gly Leu Gly Trp Gly Gly Arg Gly Ala 75 80 85 gcc gtc tgc gcc tac gtg cgc atg gtt ttc ctg gcg ctc tac gtg ctg 341 Ala Val Cys Ala Tyr Val Arg Met Val Phe Leu Ala Leu Tyr Val Leu 90 95 100 ttc ctc gcc gac gag gag ttc gac gtg gta gtg tgc gac cag gtg tct 389 Phe Leu Ala Asp Glu Glu Phe Asp Val Val Val Cys Asp Gln Val Ser 105 110 115 gcc tgt atc cca gtg ttc agg ctg gct aga cgg cgg aag aag atc cta 437 Ala Cys Ile Pro Val Phe Arg Leu Ala Arg Arg Arg Lys Lys Ile Leu 120 125 130 ttt tac tgt cac ttc cca gat ctg ctt ctc acc aag aga gat tct ttt 485 Phe Tyr Cys His Phe Pro Asp Leu Leu Leu Thr Lys Arg Asp Ser Phe 135 140 145 150 ctt aaa cga cta tac agg gcc cca att gac tgg ata gag gaa tac acc 533 Leu Lys Arg Leu Tyr Arg Ala Pro Ile Asp Trp Ile Glu Glu Tyr Thr 155 160 165 aca ggc atg gca gac tgc atc tta gtc aac agc cag ttc aca gct gct 581 Thr Gly Met Ala Asp Cys Ile Leu Val Asn Ser Gln Phe Thr Ala Ala 170 175 180 gtt ttt aag gaa aca ttc aag tcc ctg tct cac ata gac cct gat gtc 629 Val Phe Lys Glu Thr Phe Lys Ser Leu Ser His Ile Asp Pro Asp Val 185 190 195 ctc tat cca tct cta aat gtc acc agc ttt gac tca gtt gtt cct gaa 677 Leu Tyr Pro Ser Leu Asn Val Thr Ser Phe Asp Ser Val Val Pro Glu 200 205 210 aag ctg gat gac cta gtc ccc aag ggg aaa aaa ttc ctg ctg ctc tcc 725 Lys Leu Asp Asp Leu Val Pro Lys Gly Lys Lys Phe Leu Leu Leu Ser 215 220 225 230 atc aac aga tac gaa agg aag aaa aat ctg act ttg gca ctg gaa gcc 773 Ile Asn Arg Tyr Glu Arg Lys Lys Asn Leu Thr Leu Ala Leu Glu Ala 235 240 245 cta gta cag ctg cgt gga aga ttg aca tcc caa gat tgg gag agg gtt 821 Leu Val Gln Leu Arg Gly Arg Leu Thr Ser Gln Asp Trp Glu Arg Val 250 255 260 cat ctg atc gtg gca ggt ggt tat gac gag aga gtc ctg gag aat gtg 869 His Leu Ile Val Ala Gly Gly Tyr Asp Glu Arg Val Leu Glu Asn Val 265 270 275 gaa cat tat cag gaa ttg aag aaa atg gtc caa cag tcc gac ctt ggc 917 Glu His Tyr Gln Glu Leu Lys Lys Met Val Gln Gln Ser Asp Leu Gly 280 285 290 cag tat gtg acc ttc ttg agg tct ttc tca gac aaa cag aaa atc tcc 965 Gln Tyr Val Thr Phe Leu Arg Ser Phe Ser Asp Lys Gln Lys Ile Ser 295 300 305 310 ctc ctc cac agc tgc acg tgt gtg ctt tac aca cca agc aat gag cac 1013 Leu Leu His Ser Cys Thr Cys Val Leu Tyr Thr Pro Ser Asn Glu His 315 320 325 ttt ggc att gtc cct ctg gaa gcc atg tac atg cag tgc cca gtc att 1061 Phe Gly Ile Val Pro Leu Glu Ala Met Tyr Met Gln Cys Pro Val Ile 330 335 340 gct gtt aat tcg ggt gga ccc ttg gag tcc att gac cac agt gtc aca 1109 Ala Val Asn Ser Gly Gly Pro Leu Glu Ser Ile Asp His Ser Val Thr 345 350 355 ggg ttt ctg tgt gag cct gac ccg gtg cac ttc tca gaa gca ata gaa 1157 Gly Phe Leu Cys Glu Pro Asp Pro Val His Phe Ser Glu Ala Ile Glu 360 365 370 aag ttc atc cgt gaa cct tcc tta aaa gcc acc atg ggc ctg gct gga 1205 Lys Phe Ile Arg Glu Pro Ser Leu Lys Ala Thr Met Gly Leu Ala Gly 375 380 385 390 aga gcc aga gtg aag gaa aaa ttt tcc cct gaa gca ttt aca gaa cag 1253 Arg Ala Arg Val Lys Glu Lys Phe Ser Pro Glu Ala Phe Thr Glu Gln 395 400 405 ctc tac cga tat gtt acc aaa ctg ctg gta taa ctcgagtcta gagggcccgt 1306 Leu Tyr Arg Tyr Val Thr Lys Leu Leu Val * 410 415 ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc 1366 ctcccccgtg ccttgccttg accctggaag gtgccacgcc ca 1408 5 416 PRT Homo sapiens 5 Met Ala Glu Glu Gln Ala Arg Glu Arg Asp Ser Val Pro Lys Pro Ser 1 5 10 15 Val Leu Phe Leu His Pro Asp Leu Gly Val Gly Gly Ala Glu Arg Leu 20 25 30 Val Leu Asp Ala Ala Leu Ala Leu Gln Ala Arg Gly Cys Ser Val Lys 35 40 45 Ile Trp Thr Ala His Tyr Asp Pro Gly His Cys Phe Ala Glu Ser Arg 50 55 60 Glu Leu Pro Val Arg Cys Ala Gly Asp Trp Leu Pro Arg Gly Leu Gly 65 70 75 80 Trp Gly Gly Arg Gly Ala Ala Val Cys Ala Tyr Val Arg Met Val Phe 85 90 95 Leu Ala Leu Tyr Val Leu Phe Leu Ala Asp Glu Glu Phe Asp Val Val 100 105 110 Val Cys Asp Gln Val Ser Ala Cys Ile Pro Val Phe Arg Leu Ala Arg 115 120 125 Arg Arg Lys Lys Ile Leu Phe Tyr Cys His Phe Pro Asp Leu Leu Leu 130 135 140 Thr Lys Arg Asp Ser Phe Leu Lys Arg Leu Tyr Arg Ala Pro Ile Asp 145 150 155 160 Trp Ile Glu Glu Tyr Thr Thr Gly Met Ala Asp Cys Ile Leu Val Asn 165 170 175 Ser Gln Phe Thr Ala Ala Val Phe Lys Glu Thr Phe Lys Ser Leu Ser 180 185 190 His Ile Asp Pro Asp Val Leu Tyr Pro Ser Leu Asn Val Thr Ser Phe 195 200 205 Asp Ser Val Val Pro Glu Lys Leu Asp Asp Leu Val Pro Lys Gly Lys 210 215 220 Lys Phe Leu Leu Leu Ser Ile Asn Arg Tyr Glu Arg Lys Lys Asn Leu 225 230 235 240 Thr Leu Ala Leu Glu Ala Leu Val Gln Leu Arg Gly Arg Leu Thr Ser 245 250 255 Gln Asp Trp Glu Arg Val His Leu Ile Val Ala Gly Gly Tyr Asp Glu 260 265 270 Arg Val Leu Glu Asn Val Glu His Tyr Gln Glu Leu Lys Lys Met Val 275 280 285 Gln Gln Ser Asp Leu Gly Gln Tyr Val Thr Phe Leu Arg Ser Phe Ser 290 295 300 Asp Lys Gln Lys Ile Ser Leu Leu His Ser Cys Thr Cys Val Leu Tyr 305 310 315 320 Thr Pro Ser Asn Glu His Phe Gly Ile Val Pro Leu Glu Ala Met Tyr 325 330 335 Met Gln Cys Pro Val Ile Ala Val Asn Ser Gly Gly Pro Leu Glu Ser 340 345 350 Ile Asp His Ser Val Thr Gly Phe Leu Cys Glu Pro Asp Pro Val His 355 360 365 Phe Ser Glu Ala Ile Glu Lys Phe Ile Arg Glu Pro Ser Leu Lys Ala 370 375 380 Thr Met Gly Leu Ala Gly Arg Ala Arg Val Lys Glu Lys Phe Ser Pro 385 390 395 400 Glu Ala Phe Thr Glu Gln Leu Tyr Arg Tyr Val Thr Lys Leu Leu Val 405 410 415 6 1251 DNA Homo sapiens CDS (1)...(1251) 6 atg gcg gag gag cag gcc cgg gaa cgg gac tcg gtt ccc aag ccg tcg 48 Met Ala Glu Glu Gln Ala Arg Glu Arg Asp Ser Val Pro Lys Pro Ser 1 5 10 15 gtg ctg ttc ctc cac cca gac ctg ggc gtg ggc ggc gct gag cgg ctg 96 Val Leu Phe Leu His Pro Asp Leu Gly Val Gly Gly Ala Glu Arg Leu 20 25 30 gtg ttg gac gcg gcg ctg gcg ctg cag gcg cgc ggg tgt agc gtg aag 144 Val Leu Asp Ala Ala Leu Ala Leu Gln Ala Arg Gly Cys Ser Val Lys 35 40 45 atc tgg aca gcg cac tac gac ccg ggc cac tgt ttc gcc gag agc cgc 192 Ile Trp Thr Ala His Tyr Asp Pro Gly His Cys Phe Ala Glu Ser Arg 50 55 60 gag cta ccg gtg cgc tgt gcc ggg gac tgg ctg ccg cga ggc ctg ggc 240 Glu Leu Pro Val Arg Cys Ala Gly Asp Trp Leu Pro Arg Gly Leu Gly 65 70 75 80 tgg ggc ggc cgc ggc gcc gcc gtc tgc gcc tac gtg cgc atg gtt ttc 288 Trp Gly Gly Arg Gly Ala Ala Val Cys Ala Tyr Val Arg Met Val Phe 85 90 95 ctg gcg ctc tac gtg ctg ttc ctc gcc gac gag gag ttc gac gtg gta 336 Leu Ala Leu Tyr Val Leu Phe Leu Ala Asp Glu Glu Phe Asp Val Val 100 105 110 gtg tgc gac cag gtg tct gcc tgt atc cca gtg ttc agg ctg gct aga 384 Val Cys Asp Gln Val Ser Ala Cys Ile Pro Val Phe Arg Leu Ala Arg 115 120 125 cgg cgg aag aag atc cta ttt tac tgt cac ttc cca gat ctg ctt ctc 432 Arg Arg Lys Lys Ile Leu Phe Tyr Cys His Phe Pro Asp Leu Leu Leu 130 135 140 acc aag aga gat tct ttt ctt aaa cga cta tac agg gcc cca att gac 480 Thr Lys Arg Asp Ser Phe Leu Lys Arg Leu Tyr Arg Ala Pro Ile Asp 145 150 155 160 tgg ata gag gaa tac acc aca ggc atg gca gac tgc atc tta gtc aac 528 Trp Ile Glu Glu Tyr Thr Thr Gly Met Ala Asp Cys Ile Leu Val Asn 165 170 175 agc cag ttc aca gct gct gtt ttt aag gaa aca ttc aag tcc ctg tct 576 Ser Gln Phe Thr Ala Ala Val Phe Lys Glu Thr Phe Lys Ser Leu Ser 180 185 190 cac ata gac cct gat gtc ctc tat cca tct cta aat gtc acc agc ttt 624 His Ile Asp Pro Asp Val Leu Tyr Pro Ser Leu Asn Val Thr Ser Phe 195 200 205 gac tca gtt gtt cct gaa aag ctg gat gac cta gtc ccc aag ggg aaa 672 Asp Ser Val Val Pro Glu Lys Leu Asp Asp Leu Val Pro Lys Gly Lys 210 215 220 aaa ttc ctg ctg ctc tcc atc aac aga tac gaa agg aag aaa aat ctg 720 Lys Phe Leu Leu Leu Ser Ile Asn Arg Tyr Glu Arg Lys Lys Asn Leu 225 230 235 240 act ttg gca ctg gaa gcc cta gta cag ctg cgt gga aga ttg aca tcc 768 Thr Leu Ala Leu Glu Ala Leu Val Gln Leu Arg Gly Arg Leu Thr Ser 245 250 255 caa gat tgg gag agg gtt cat ctg atc gtg gca ggt ggt tat gac gag 816 Gln Asp Trp Glu Arg Val His Leu Ile Val Ala Gly Gly Tyr Asp Glu 260 265 270 aga gtc ctg gag aat gtg gaa cat tat cag gaa ttg aag aaa atg gtc 864 Arg Val Leu Glu Asn Val Glu His Tyr Gln Glu Leu Lys Lys Met Val 275 280 285 caa cag tcc gac ctt ggc cag tat gtg acc ttc ttg agg tct ttc tca 912 Gln Gln Ser Asp Leu Gly Gln Tyr Val Thr Phe Leu Arg Ser Phe Ser 290 295 300 gac aaa cag aaa atc tcc ctc ctc cac agc tgc acg tgt gtg ctt tac 960 Asp Lys Gln Lys Ile Ser Leu Leu His Ser Cys Thr Cys Val Leu Tyr 305 310 315 320 aca cca agc aat gag cac ttt ggc att gtc cct ctg gaa gcc atg tac 1008 Thr Pro Ser Asn Glu His Phe Gly Ile Val Pro Leu Glu Ala Met Tyr 325 330 335 atg cag tgc cca gtc att gct gtt aat tcg ggt gga ccc ttg gag tcc 1056 Met Gln Cys Pro Val Ile Ala Val Asn Ser Gly Gly Pro Leu Glu Ser 340 345 350 att gac cac agt gtc aca ggg ttt ctg tgt gag cct gac ccg gtg cac 1104 Ile Asp His Ser Val Thr Gly Phe Leu Cys Glu Pro Asp Pro Val His 355 360 365 ttc tca gaa gca ata gaa aag ttc atc cgt gaa cct tcc tta aaa gcc 1152 Phe Ser Glu Ala Ile Glu Lys Phe Ile Arg Glu Pro Ser Leu Lys Ala 370 375 380 acc atg ggc ctg gct gga aga gcc aga gtg aag gaa aaa ttt tcc cct 1200 Thr Met Gly Leu Ala Gly Arg Ala Arg Val Lys Glu Lys Phe Ser Pro 385 390 395 400 gaa gca ttt aca gaa cag ctc tac cga tat gtt acc aaa ctg ctg gta 1248 Glu Ala Phe Thr Glu Gln Leu Tyr Arg Tyr Val Thr Lys Leu Leu Val 405 410 415 taa 1251 * 7 2327 DNA Homo sapiens CDS (148)...(1974) 7 ggcagttcag cccgcgccgc tcctgcgggt cggactgggg ctgtggcggg agagaagatg 60 ccgcagcccg agtcccagaa ggcggcgatc ctgggctgcg ggcaaggcgg aaattgacaa 120 tggcccttca gctatgctag gtctata atg gga agt gtc aca gtt cgg tat ttc 174 Met Gly Ser Val Thr Val Arg Tyr Phe 1 5 tgt tat ggg tgc ctt ttt aca tct gcg acc tgg aca gtt ttg ctt ttt 222 Cys Tyr Gly Cys Leu Phe Thr Ser Ala Thr Trp Thr Val Leu Leu Phe 10 15 20 25 gtt tat ttc aac ttc agt gaa gtg act cag cca ctt aag aat gtg ccc 270 Val Tyr Phe Asn Phe Ser Glu Val Thr Gln Pro Leu Lys Asn Val Pro 30 35 40 gtc aag ggg tct ggg ccc cac gga cca tct cca aaa aaa ttc tat ccc 318 Val Lys Gly Ser Gly Pro His Gly Pro Ser Pro Lys Lys Phe Tyr Pro 45 50 55 cgt ttc act cga ggc cca agt cga gtg ctc gag cca cag ttc aaa gca 366 Arg Phe Thr Arg Gly Pro Ser Arg Val Leu Glu Pro Gln Phe Lys Ala 60 65 70 aac aaa att gac gat gtg ata gac agt cgt gtt gaa gat cca gaa gaa 414 Asn Lys Ile Asp Asp Val Ile Asp Ser Arg Val Glu Asp Pro Glu Glu 75 80 85 ggc cac ttg aaa ttc tct tct gaa tta ggt atg att ttt aat gaa cgc 462 Gly His Leu Lys Phe Ser Ser Glu Leu Gly Met Ile Phe Asn Glu Arg 90 95 100 105 gat caa gag ttg aga gac ttg ggc tat cag aaa cat gct ttt aat atg 510 Asp Gln Glu Leu Arg Asp Leu Gly Tyr Gln Lys His Ala Phe Asn Met 110 115 120 ctt atc agt gac cgc ttg ggc tac cac aga gat gtg cca gac aca agg 558 Leu Ile Ser Asp Arg Leu Gly Tyr His Arg Asp Val Pro Asp Thr Arg 125 130 135 aat gca gca tgt aaa gaa aag ttc tac cca cct gac ctg cca gct gct 606 Asn Ala Ala Cys Lys Glu Lys Phe Tyr Pro Pro Asp Leu Pro Ala Ala 140 145 150 agt gtt gtt atc tgt ttc tat aat gaa gcg ttt tct gcc ttg ctt cgg 654 Ser Val Val Ile Cys Phe Tyr Asn Glu Ala Phe Ser Ala Leu Leu Arg 155 160 165 aca gtg cac agt gtc ata gac cgc acg cca gca cac ctg ctt cat gag 702 Thr Val His Ser Val Ile Asp Arg Thr Pro Ala His Leu Leu His Glu 170 175 180 185 atc atc ctt gtg gat gat gat agt gac ttt gat gat ttg aaa gga gaa 750 Ile Ile Leu Val Asp Asp Asp Ser Asp Phe Asp Asp Leu Lys Gly Glu 190 195 200 cta gat gaa tat gtc caa aaa tac ctc cct gga aaa att aaa gtc ata 798 Leu Asp Glu Tyr Val Gln Lys Tyr Leu Pro Gly Lys Ile Lys Val Ile 205 210 215 aga aat aca aag cgt gag ggg ttg att cga ggg aga atg att ggc gcg 846 Arg Asn Thr Lys Arg Glu Gly Leu Ile Arg Gly Arg Met Ile Gly Ala 220 225 230 gcc cac gcg aca gga gaa gtc ctt gtg ttc ctg gac agc cac tgt gaa 894 Ala His Ala Thr Gly Glu Val Leu Val Phe Leu Asp Ser His Cys Glu 235 240 245 gtg aat gtg atg tgg ctg cag ccc ttg ctg gcc gcc atc cgt gag gac 942 Val Asn Val Met Trp Leu Gln Pro Leu Leu Ala Ala Ile Arg Glu Asp 250 255 260 265 cgg cac acc gtg gtg tgc cca gtg att gac atc atc agc gcc gac acg 990 Arg His Thr Val Val Cys Pro Val Ile Asp Ile Ile Ser Ala Asp Thr 270 275 280 ctg gcc tac agc tcg tcc cct gtc gtc cgc gga ggg ttc aac tgg gga 1038 Leu Ala Tyr Ser Ser Ser Pro Val Val Arg Gly Gly Phe Asn Trp Gly 285 290 295 ctg cac ttc aaa tgg gat ctt gtc ccc ctt tct gag cta gga cga gcg 1086 Leu His Phe Lys Trp Asp Leu Val Pro Leu Ser Glu Leu Gly Arg Ala 300 305 310 gag gga gcc act gca cca ata aag tca cca aca atg gct gga ggt ttg 1134 Glu Gly Ala Thr Ala Pro Ile Lys Ser Pro Thr Met Ala Gly Gly Leu 315 320 325 ttt gcc atg aac aga cag tat ttc cat gaa ctt gga cag tat gat agt 1182 Phe Ala Met Asn Arg Gln Tyr Phe His Glu Leu Gly Gln Tyr Asp Ser 330 335 340 345 ggc atg gat atc tgg gga gga gaa aat ttg gaa ata tca ttt cgg atc 1230 Gly Met Asp Ile Trp Gly Gly Glu Asn Leu Glu Ile Ser Phe Arg Ile 350 355 360 tgg atg tgt ggc ggt aag ctc ttc atc atc cct tgc tct aga gta gga 1278 Trp Met Cys Gly Gly Lys Leu Phe Ile Ile Pro Cys Ser Arg Val Gly 365 370 375 cac att ttc cga aaa agg cga cca tat gga tct ccc gaa ggc cag gac 1326 His Ile Phe Arg Lys Arg Arg Pro Tyr Gly Ser Pro Glu Gly Gln Asp 380 385 390 acc atg aca cac aac tct ttg cgg ctg gca cat gtc tgg ttg gat gaa 1374 Thr Met Thr His Asn Ser Leu Arg Leu Ala His Val Trp Leu Asp Glu 395 400 405 tac aag gag cag tat ttt tcc tta aga cct gac ctg aag acg aaa agc 1422 Tyr Lys Glu Gln Tyr Phe Ser Leu Arg Pro Asp Leu Lys Thr Lys Ser 410 415 420 425 tat ggc aat atc agt gag cgt gtg gaa ctg aga aag aag ttg ggc tgt 1470 Tyr Gly Asn Ile Ser Glu Arg Val Glu Leu Arg Lys Lys Leu Gly Cys 430 435 440 aaa tca ttt aaa tgg tat ttg gat aat gta tac cca gag atg cag ata 1518 Lys Ser Phe Lys Trp Tyr Leu Asp Asn Val Tyr Pro Glu Met Gln Ile 445 450 455 tct ggg tcc cac gcc aaa ccc caa caa ccc att ttt gtc aat aga ggg 1566 Ser Gly Ser His Ala Lys Pro Gln Gln Pro Ile Phe Val Asn Arg Gly 460 465 470 cca aaa cga ccc aaa gtc ctt caa cgt gga agg ctc tat cac ctc cag 1614 Pro Lys Arg Pro Lys Val Leu Gln Arg Gly Arg Leu Tyr His Leu Gln 475 480 485 acc aac aaa tgc ctg gtg gcc cag ggc cgc cca agt cag aag gga ggt 1662 Thr Asn Lys Cys Leu Val Ala Gln Gly Arg Pro Ser Gln Lys Gly Gly 490 495 500 505 ctc gtg gtg ctt aag gcc tgt gac tac agt gac cca aat cag atc tgg 1710 Leu Val Val Leu Lys Ala Cys Asp Tyr Ser Asp Pro Asn Gln Ile Trp 510 515 520 atc tat aat gaa gag cat gaa ttg gtt tta aat agt ctc ctt tgt cta 1758 Ile Tyr Asn Glu Glu His Glu Leu Val Leu Asn Ser Leu Leu Cys Leu 525 530 535 gat atg tca gag act cgc tca tca gac ccg cca cgg ctc atg aaa tgc 1806 Asp Met Ser Glu Thr Arg Ser Ser Asp Pro Pro Arg Leu Met Lys Cys 540 545 550 cac ggg tca gga gga tcc cag cag tgg acc ttt ggg aaa aac aat cgg 1854 His Gly Ser Gly Gly Ser Gln Gln Trp Thr Phe Gly Lys Asn Asn Arg 555 560 565 cta tac cag gtg tcg gtt gga cag tgc ctg aga gca gtg gat ccc ctg 1902 Leu Tyr Gln Val Ser Val Gly Gln Cys Leu Arg Ala Val Asp Pro Leu 570 575 580 585 ggt cag aag ggc tct gtc gcc atg gcg atc tgc gat ggc tcc tct tca 1950 Gly Gln Lys Gly Ser Val Ala Met Ala Ile Cys Asp Gly Ser Ser Ser 590 595 600 cag cag tgg cat ttg gaa ggt taa ggtggatgct gtggcgggaa cgttgcttca 2004 Gln Gln Trp His Leu Glu Gly * 605 tcaggcgttg cctccggtgt ggagtttggg gctttaggaa agcctgggtt gggtggagca 2064 gaaccatctt ggagaagatg acagttccct gtcctcccgg agatgcctgg gtgtgttagc 2124 agaggtgaca cgtgtctgac agagacggga gctctgagtg tccacgggtg aagaagtgag 2184 tgtccacggg tgaagaagtg agtatgtttc acctggacat taaggtgatg tttgagctgc 2244 tgttaaggaa tttcttgctt atagaggcaa accacagtat cattttaact ctagaattgg 2304 gcttgtacct cggccgcgac cac 2327 8 608 PRT Homo sapiens 8 Met Gly Ser Val Thr Val Arg Tyr Phe Cys Tyr Gly Cys Leu Phe Thr 1 5 10 15 Ser Ala Thr Trp Thr Val Leu Leu Phe Val Tyr Phe Asn Phe Ser Glu 20 25 30 Val Thr Gln Pro Leu Lys Asn Val Pro Val Lys Gly Ser Gly Pro His 35 40 45 Gly Pro Ser Pro Lys Lys Phe Tyr Pro Arg Phe Thr Arg Gly Pro Ser 50 55 60 Arg Val Leu Glu Pro Gln Phe Lys Ala Asn Lys Ile Asp Asp Val Ile 65 70 75 80 Asp Ser Arg Val Glu Asp Pro Glu Glu Gly His Leu Lys Phe Ser Ser 85 90 95 Glu Leu Gly Met Ile Phe Asn Glu Arg Asp Gln Glu Leu Arg Asp Leu 100 105 110 Gly Tyr Gln Lys His Ala Phe Asn Met Leu Ile Ser Asp Arg Leu Gly 115 120 125 Tyr His Arg Asp Val Pro Asp Thr Arg Asn Ala Ala Cys Lys Glu Lys 130 135 140 Phe Tyr Pro Pro Asp Leu Pro Ala Ala Ser Val Val Ile Cys Phe Tyr 145 150 155 160 Asn Glu Ala Phe Ser Ala Leu Leu Arg Thr Val His Ser Val Ile Asp 165 170 175 Arg Thr Pro Ala His Leu Leu His Glu Ile Ile Leu Val Asp Asp Asp 180 185 190 Ser Asp Phe Asp Asp Leu Lys Gly Glu Leu Asp Glu Tyr Val Gln Lys 195 200 205 Tyr Leu Pro Gly Lys Ile Lys Val Ile Arg Asn Thr Lys Arg Glu Gly 210 215 220 Leu Ile Arg Gly Arg Met Ile Gly Ala Ala His Ala Thr Gly Glu Val 225 230 235 240 Leu Val Phe Leu Asp Ser His Cys Glu Val Asn Val Met Trp Leu Gln 245 250 255 Pro Leu Leu Ala Ala Ile Arg Glu Asp Arg His Thr Val Val Cys Pro 260 265 270 Val Ile Asp Ile Ile Ser Ala Asp Thr Leu Ala Tyr Ser Ser Ser Pro 275 280 285 Val Val Arg Gly Gly Phe Asn Trp Gly Leu His Phe Lys Trp Asp Leu 290 295 300 Val Pro Leu Ser Glu Leu Gly Arg Ala Glu Gly Ala Thr Ala Pro Ile 305 310 315 320 Lys Ser Pro Thr Met Ala Gly Gly Leu Phe Ala Met Asn Arg Gln Tyr 325 330 335 Phe His Glu Leu Gly Gln Tyr Asp Ser Gly Met Asp Ile Trp Gly Gly 340 345 350 Glu Asn Leu Glu Ile Ser Phe Arg Ile Trp Met Cys Gly Gly Lys Leu 355 360 365 Phe Ile Ile Pro Cys Ser Arg Val Gly His Ile Phe Arg Lys Arg Arg 370 375 380 Pro Tyr Gly Ser Pro Glu Gly Gln Asp Thr Met Thr His Asn Ser Leu 385 390 395 400 Arg Leu Ala His Val Trp Leu Asp Glu Tyr Lys Glu Gln Tyr Phe Ser 405 410 415 Leu Arg Pro Asp Leu Lys Thr Lys Ser Tyr Gly Asn Ile Ser Glu Arg 420 425 430 Val Glu Leu Arg Lys Lys Leu Gly Cys Lys Ser Phe Lys Trp Tyr Leu 435 440 445 Asp Asn Val Tyr Pro Glu Met Gln Ile Ser Gly Ser His Ala Lys Pro 450 455 460 Gln Gln Pro Ile Phe Val Asn Arg Gly Pro Lys Arg Pro Lys Val Leu 465 470 475 480 Gln Arg Gly Arg Leu Tyr His Leu Gln Thr Asn Lys Cys Leu Val Ala 485 490 495 Gln Gly Arg Pro Ser Gln Lys Gly Gly Leu Val Val Leu Lys Ala Cys 500 505 510 Asp Tyr Ser Asp Pro Asn Gln Ile Trp Ile Tyr Asn Glu Glu His Glu 515 520 525 Leu Val Leu Asn Ser Leu Leu Cys Leu Asp Met Ser Glu Thr Arg Ser 530 535 540 Ser Asp Pro Pro Arg Leu Met Lys Cys His Gly Ser Gly Gly Ser Gln 545 550 555 560 Gln Trp Thr Phe Gly Lys Asn Asn Arg Leu Tyr Gln Val Ser Val Gly 565 570 575 Gln Cys Leu Arg Ala Val Asp Pro Leu Gly Gln Lys Gly Ser Val Ala 580 585 590 Met Ala Ile Cys Asp Gly Ser Ser Ser Gln Gln Trp His Leu Glu Gly 595 600 605 9 1827 DNA Homo sapiens CDS (1)...(1827) 9 atg gga agt gtc aca gtt cgg tat ttc tgt tat ggg tgc ctt ttt aca 48 Met Gly Ser Val Thr Val Arg Tyr Phe Cys Tyr Gly Cys Leu Phe Thr 1 5 10 15 tct gcg acc tgg aca gtt ttg ctt ttt gtt tat ttc aac ttc agt gaa 96 Ser Ala Thr Trp Thr Val Leu Leu Phe Val Tyr Phe Asn Phe Ser Glu 20 25 30 gtg act cag cca ctt aag aat gtg ccc gtc aag ggg tct ggg ccc cac 144 Val Thr Gln Pro Leu Lys Asn Val Pro Val Lys Gly Ser Gly Pro His 35 40 45 gga cca tct cca aaa aaa ttc tat ccc cgt ttc act cga ggc cca agt 192 Gly Pro Ser Pro Lys Lys Phe Tyr Pro Arg Phe Thr Arg Gly Pro Ser 50 55 60 cga gtg ctc gag cca cag ttc aaa gca aac aaa att gac gat gtg ata 240 Arg Val Leu Glu Pro Gln Phe Lys Ala Asn Lys Ile Asp Asp Val Ile 65 70 75 80 gac agt cgt gtt gaa gat cca gaa gaa ggc cac ttg aaa ttc tct tct 288 Asp Ser Arg Val Glu Asp Pro Glu Glu Gly His Leu Lys Phe Ser Ser 85 90 95 gaa tta ggt atg att ttt aat gaa cgc gat caa gag ttg aga gac ttg 336 Glu Leu Gly Met Ile Phe Asn Glu Arg Asp Gln Glu Leu Arg Asp Leu 100 105 110 ggc tat cag aaa cat gct ttt aat atg ctt atc agt gac cgc ttg ggc 384 Gly Tyr Gln Lys His Ala Phe Asn Met Leu Ile Ser Asp Arg Leu Gly 115 120 125 tac cac aga gat gtg cca gac aca agg aat gca gca tgt aaa gaa aag 432 Tyr His Arg Asp Val Pro Asp Thr Arg Asn Ala Ala Cys Lys Glu Lys 130 135 140 ttc tac cca cct gac ctg cca gct gct agt gtt gtt atc tgt ttc tat 480 Phe Tyr Pro Pro Asp Leu Pro Ala Ala Ser Val Val Ile Cys Phe Tyr 145 150 155 160 aat gaa gcg ttt tct gcc ttg ctt cgg aca gtg cac agt gtc ata gac 528 Asn Glu Ala Phe Ser Ala Leu Leu Arg Thr Val His Ser Val Ile Asp 165 170 175 cgc acg cca gca cac ctg ctt cat gag atc atc ctt gtg gat gat gat 576 Arg Thr Pro Ala His Leu Leu His Glu Ile Ile Leu Val Asp Asp Asp 180 185 190 agt gac ttt gat gat ttg aaa gga gaa cta gat gaa tat gtc caa aaa 624 Ser Asp Phe Asp Asp Leu Lys Gly Glu Leu Asp Glu Tyr Val Gln Lys 195 200 205 tac ctc cct gga aaa att aaa gtc ata aga aat aca aag cgt gag ggg 672 Tyr Leu Pro Gly Lys Ile Lys Val Ile Arg Asn Thr Lys Arg Glu Gly 210 215 220 ttg att cga ggg aga atg att ggc gcg gcc cac gcg aca gga gaa gtc 720 Leu Ile Arg Gly Arg Met Ile Gly Ala Ala His Ala Thr Gly Glu Val 225 230 235 240 ctt gtg ttc ctg gac agc cac tgt gaa gtg aat gtg atg tgg ctg cag 768 Leu Val Phe Leu Asp Ser His Cys Glu Val Asn Val Met Trp Leu Gln 245 250 255 ccc ttg ctg gcc gcc atc cgt gag gac cgg cac acc gtg gtg tgc cca 816 Pro Leu Leu Ala Ala Ile Arg Glu Asp Arg His Thr Val Val Cys Pro 260 265 270 gtg att gac atc atc agc gcc gac acg ctg gcc tac agc tcg tcc cct 864 Val Ile Asp Ile Ile Ser Ala Asp Thr Leu Ala Tyr Ser Ser Ser Pro 275 280 285 gtc gtc cgc gga ggg ttc aac tgg gga ctg cac ttc aaa tgg gat ctt 912 Val Val Arg Gly Gly Phe Asn Trp Gly Leu His Phe Lys Trp Asp Leu 290 295 300 gtc ccc ctt tct gag cta gga cga gcg gag gga gcc act gca cca ata 960 Val Pro Leu Ser Glu Leu Gly Arg Ala Glu Gly Ala Thr Ala Pro Ile 305 310 315 320 aag tca cca aca atg gct gga ggt ttg ttt gcc atg aac aga cag tat 1008 Lys Ser Pro Thr Met Ala Gly Gly Leu Phe Ala Met Asn Arg Gln Tyr 325 330 335 ttc cat gaa ctt gga cag tat gat agt ggc atg gat atc tgg gga gga 1056 Phe His Glu Leu Gly Gln Tyr Asp Ser Gly Met Asp Ile Trp Gly Gly 340 345 350 gaa aat ttg gaa ata tca ttt cgg atc tgg atg tgt ggc ggt aag ctc 1104 Glu Asn Leu Glu Ile Ser Phe Arg Ile Trp Met Cys Gly Gly Lys Leu 355 360 365 ttc atc atc cct tgc tct aga gta gga cac att ttc cga aaa agg cga 1152 Phe Ile Ile Pro Cys Ser Arg Val Gly His Ile Phe Arg Lys Arg Arg 370 375 380 cca tat gga tct ccc gaa ggc cag gac acc atg aca cac aac tct ttg 1200 Pro Tyr Gly Ser Pro Glu Gly Gln Asp Thr Met Thr His Asn Ser Leu 385 390 395 400 cgg ctg gca cat gtc tgg ttg gat gaa tac aag gag cag tat ttt tcc 1248 Arg Leu Ala His Val Trp Leu Asp Glu Tyr Lys Glu Gln Tyr Phe Ser 405 410 415 tta aga cct gac ctg aag acg aaa agc tat ggc aat atc agt gag cgt 1296 Leu Arg Pro Asp Leu Lys Thr Lys Ser Tyr Gly Asn Ile Ser Glu Arg 420 425 430 gtg gaa ctg aga aag aag ttg ggc tgt aaa tca ttt aaa tgg tat ttg 1344 Val Glu Leu Arg Lys Lys Leu Gly Cys Lys Ser Phe Lys Trp Tyr Leu 435 440 445 gat aat gta tac cca gag atg cag ata tct ggg tcc cac gcc aaa ccc 1392 Asp Asn Val Tyr Pro Glu Met Gln Ile Ser Gly Ser His Ala Lys Pro 450 455 460 caa caa ccc att ttt gtc aat aga ggg cca aaa cga ccc aaa gtc ctt 1440 Gln Gln Pro Ile Phe Val Asn Arg Gly Pro Lys Arg Pro Lys Val Leu 465 470 475 480 caa cgt gga agg ctc tat cac ctc cag acc aac aaa tgc ctg gtg gcc 1488 Gln Arg Gly Arg Leu Tyr His Leu Gln Thr Asn Lys Cys Leu Val Ala 485 490 495 cag ggc cgc cca agt cag aag gga ggt ctc gtg gtg ctt aag gcc tgt 1536 Gln Gly Arg Pro Ser Gln Lys Gly Gly Leu Val Val Leu Lys Ala Cys 500 505 510 gac tac agt gac cca aat cag atc tgg atc tat aat gaa gag cat gaa 1584 Asp Tyr Ser Asp Pro Asn Gln Ile Trp Ile Tyr Asn Glu Glu His Glu 515 520 525 ttg gtt tta aat agt ctc ctt tgt cta gat atg tca gag act cgc tca 1632 Leu Val Leu Asn Ser Leu Leu Cys Leu Asp Met Ser Glu Thr Arg Ser 530 535 540 tca gac ccg cca cgg ctc atg aaa tgc cac ggg tca gga gga tcc cag 1680 Ser Asp Pro Pro Arg Leu Met Lys Cys His Gly Ser Gly Gly Ser Gln 545 550 555 560 cag tgg acc ttt ggg aaa aac aat cgg cta tac cag gtg tcg gtt gga 1728 Gln Trp Thr Phe Gly Lys Asn Asn Arg Leu Tyr Gln Val Ser Val Gly 565 570 575 cag tgc ctg aga gca gtg gat ccc ctg ggt cag aag ggc tct gtc gcc 1776 Gln Cys Leu Arg Ala Val Asp Pro Leu Gly Gln Lys Gly Ser Val Ala 580 585 590 atg gcg atc tgc gat ggc tcc tct tca cag cag tgg cat ttg gaa ggt 1824 Met Ala Ile Cys Asp Gly Ser Ser Ser Gln Gln Trp His Leu Glu Gly 595 600 605 taa 1827 * 10 2172 DNA Homo sapiens CDS (31)...(2019) 10 gcgctgcgct gctggggggc gcgggcgagg atg gcg gcg gag aac gag gcc agc 54 Met Ala Ala Glu Asn Glu Ala Ser 1 5 cag gag agc gcc ctg ggc gcc tac tcg cca gtg gac tac atg agc atc 102 Gln Glu Ser Ala Leu Gly Ala Tyr Ser Pro Val Asp Tyr Met Ser Ile 10 15 20 acc agc ttc ccg cgg ctg ccc gag gac gag ccg gcg ccc gcg gcc ccg 150 Thr Ser Phe Pro Arg Leu Pro Glu Asp Glu Pro Ala Pro Ala Ala Pro 25 30 35 40 ctg agg ggc cgc aag gac gag gac gcc ttt ctg gga gac ccc gac acc 198 Leu Arg Gly Arg Lys Asp Glu Asp Ala Phe Leu Gly Asp Pro Asp Thr 45 50 55 gac ccg gac tcc ttc ctg aag tct gca cgg ctg cag cgg ctg cca tcg 246 Asp Pro Asp Ser Phe Leu Lys Ser Ala Arg Leu Gln Arg Leu Pro Ser 60 65 70 tcg tcg tcg gag atg ggc agc caa gac ggg tcg ccg cta cgc gag acg 294 Ser Ser Ser Glu Met Gly Ser Gln Asp Gly Ser Pro Leu Arg Glu Thr 75 80 85 cgc aaa gac ccg ttc tcc gcc gca gcg gcc gag tgc tcc tgc cgc cag 342 Arg Lys Asp Pro Phe Ser Ala Ala Ala Ala Glu Cys Ser Cys Arg Gln 90 95 100 gat ggg ctc acg gtc atc gtc acg gcc tgt ctc acc ttc gct acc ggt 390 Asp Gly Leu Thr Val Ile Val Thr Ala Cys Leu Thr Phe Ala Thr Gly 105 110 115 120 gtc acc gtg gcg ctg gtc atg cag atc tac ttc ggg gac ccc cag atc 438 Val Thr Val Ala Leu Val Met Gln Ile Tyr Phe Gly Asp Pro Gln Ile 125 130 135 ttc cag cag ggt gcc gtg gtg acc gat gct gcc cgc tgc act tca ctg 486 Phe Gln Gln Gly Ala Val Val Thr Asp Ala Ala Arg Cys Thr Ser Leu 140 145 150 ggc atc gag gtg ctc agt aaa cag gga tct tct gtg gac gca gcg gtg 534 Gly Ile Glu Val Leu Ser Lys Gln Gly Ser Ser Val Asp Ala Ala Val 155 160 165 gca gca gcc ttg tgt ttg ggt atc gtg gct cca cac agt tct ggc ctg 582 Ala Ala Ala Leu Cys Leu Gly Ile Val Ala Pro His Ser Ser Gly Leu 170 175 180 ggc ggt ggg ggc gtg atg ctg gta cat gac atc cga cga aat gag agc 630 Gly Gly Gly Gly Val Met Leu Val His Asp Ile Arg Arg Asn Glu Ser 185 190 195 200 cac cta att gat ttc cgg gag tcc gca cca ggg gcc ctc agg gaa gag 678 His Leu Ile Asp Phe Arg Glu Ser Ala Pro Gly Ala Leu Arg Glu Glu 205 210 215 acc ctg caa aga tcc tgg gag acc aag cct ggg ctc ttg gtg ggg gtt 726 Thr Leu Gln Arg Ser Trp Glu Thr Lys Pro Gly Leu Leu Val Gly Val 220 225 230 ccc gga atg gtg aag ggg cta cat gaa gct cac cag ctc tat ggc agg 774 Pro Gly Met Val Lys Gly Leu His Glu Ala His Gln Leu Tyr Gly Arg 235 240 245 ctg cca tgg tcc caa gtc ctg gcc ttt gca gca gct gtg gcc caa gat 822 Leu Pro Trp Ser Gln Val Leu Ala Phe Ala Ala Ala Val Ala Gln Asp 250 255 260 ggc ttc aac gtg act cat gat cta gcc cgt gcc ctg gct gaa cag ctg 870 Gly Phe Asn Val Thr His Asp Leu Ala Arg Ala Leu Ala Glu Gln Leu 265 270 275 280 cca ccc aac atg tcc gag cgc ttc cgg gag acg ttc ctg cca tcg ggc 918 Pro Pro Asn Met Ser Glu Arg Phe Arg Glu Thr Phe Leu Pro Ser Gly 285 290 295 cgc ccg cca cta cct ggc tcg ttg ctg cat cgg ccc gac ctg gct gag 966 Arg Pro Pro Leu Pro Gly Ser Leu Leu His Arg Pro Asp Leu Ala Glu 300 305 310 gtg ctg gat gta ctt ggc acc tcc ggc ccg gct gcc ttc tac gca ggt 1014 Val Leu Asp Val Leu Gly Thr Ser Gly Pro Ala Ala Phe Tyr Ala Gly 315 320 325 ggc aac ctc aca ctg gag atg gtg gcc gag gct cag cac gca ggg ggt 1062 Gly Asn Leu Thr Leu Glu Met Val Ala Glu Ala Gln His Ala Gly Gly 330 335 340 gtc ata acc gaa gag gac ttc agc aat tac agc gcc ctt gtg gag aag 1110 Val Ile Thr Glu Glu Asp Phe Ser Asn Tyr Ser Ala Leu Val Glu Lys 345 350 355 360 cct gtg tgt ggc gtg tac aga ggc cac ctg gtt ctt agt ccc cca cct 1158 Pro Val Cys Gly Val Tyr Arg Gly His Leu Val Leu Ser Pro Pro Pro 365 370 375 ccg cac acg ggc cct gcc ctc atc agt gct ctc aac atc ctg gag ggc 1206 Pro His Thr Gly Pro Ala Leu Ile Ser Ala Leu Asn Ile Leu Glu Gly 380 385 390 ttc aat ctc acc agc ctg gta tcc cga gaa cag gct ctt cac tgg gtg 1254 Phe Asn Leu Thr Ser Leu Val Ser Arg Glu Gln Ala Leu His Trp Val 395 400 405 gca gag acc ctg aag att gca tta gcc ctg gcc agc aga ctg gga gat 1302 Ala Glu Thr Leu Lys Ile Ala Leu Ala Leu Ala Ser Arg Leu Gly Asp 410 415 420 ccc gtc tat gat tct acc atc act gag agc atg gat gac atg ctc agc 1350 Pro Val Tyr Asp Ser Thr Ile Thr Glu Ser Met Asp Asp Met Leu Ser 425 430 435 440 aag gtg gag gcc gcc tac ctc cgg ggc cat atc aat gac tcc cag gca 1398 Lys Val Glu Ala Ala Tyr Leu Arg Gly His Ile Asn Asp Ser Gln Ala 445 450 455 gcc cct gcc cca ctc ctg cct gtc tat gaa cta gac gga gct ccc acg 1446 Ala Pro Ala Pro Leu Leu Pro Val Tyr Glu Leu Asp Gly Ala Pro Thr 460 465 470 gct gcc cag gtg ctg atc atg gga cct gat gac ttc att gtg gcc atg 1494 Ala Ala Gln Val Leu Ile Met Gly Pro Asp Asp Phe Ile Val Ala Met 475 480 485 gtt agc tcc ctg aac cag ccc ttt ggc agc ggc ctt atc acc ccc tcg 1542 Val Ser Ser Leu Asn Gln Pro Phe Gly Ser Gly Leu Ile Thr Pro Ser 490 495 500 ggg atc ctg ctc aac agc cag atg ctg gac ttc tcc tgg ccc aac cgg 1590 Gly Ile Leu Leu Asn Ser Gln Met Leu Asp Phe Ser Trp Pro Asn Arg 505 510 515 520 aca gct aac cac tct gca ccc agc ctg gag aat tca gtg cag cca ggg 1638 Thr Ala Asn His Ser Ala Pro Ser Leu Glu Asn Ser Val Gln Pro Gly 525 530 535 aag cgg cca ctc tct ttc ctg ctg ccc aca gtg gtc cga ccc gcg gag 1686 Lys Arg Pro Leu Ser Phe Leu Leu Pro Thr Val Val Arg Pro Ala Glu 540 545 550 ggg ctc tgt gga acc tac ctc gct ctg ggg gcc aat gga gct gcg cgg 1734 Gly Leu Cys Gly Thr Tyr Leu Ala Leu Gly Ala Asn Gly Ala Ala Arg 555 560 565 ggc ctc agc ggc ctg aca cag gtt ctg ctg aat gtc ctg acc ttg aac 1782 Gly Leu Ser Gly Leu Thr Gln Val Leu Leu Asn Val Leu Thr Leu Asn 570 575 580 cgg aac ctg agt gac agc ctg gcc cgc ggc cgc cta cac ccg gac ctg 1830 Arg Asn Leu Ser Asp Ser Leu Ala Arg Gly Arg Leu His Pro Asp Leu 585 590 595 600 cag tcc aac ctc ctg cag gtg gac agt gag ttc aca gag gaa gag att 1878 Gln Ser Asn Leu Leu Gln Val Asp Ser Glu Phe Thr Glu Glu Glu Ile 605 610 615 gag ttc ctg gaa gcc agg ggt cac cac gtg gag aaa gta gat gtc tta 1926 Glu Phe Leu Glu Ala Arg Gly His His Val Glu Lys Val Asp Val Leu 620 625 630 tcc tgg gtc cat ggc agc cga agg acc aac aac ttc atc atc gct gtt 1974 Ser Trp Val His Gly Ser Arg Arg Thr Asn Asn Phe Ile Ile Ala Val 635 640 645 aag gac cct cgg agc cca gat gca gct gga gcc acc atc ctg tag 2019 Lys Asp Pro Arg Ser Pro Asp Ala Ala Gly Ala Thr Ile Leu * 650 655 660 agcagcgggg tggggcgggg tctctgctcc cccactttgc atgttcccag agtccctcct 2079 tctcccaggt ttggtctcag ggggacccca gggatgcccc agatcagggg ccagagggga 2139 tgcttagcaa acccaatccc agagtaactg gaa 2172 11 662 PRT Homo sapiens 11 Met Ala Ala Glu Asn Glu Ala Ser Gln Glu Ser Ala Leu Gly Ala Tyr 1 5 10 15 Ser Pro Val Asp Tyr Met Ser Ile Thr Ser Phe Pro Arg Leu Pro Glu 20 25 30 Asp Glu Pro Ala Pro Ala Ala Pro Leu Arg Gly Arg Lys Asp Glu Asp 35 40 45 Ala Phe Leu Gly Asp Pro Asp Thr Asp Pro Asp Ser Phe Leu Lys Ser 50 55 60 Ala Arg Leu Gln Arg Leu Pro Ser Ser Ser Ser Glu Met Gly Ser Gln 65 70 75 80 Asp Gly Ser Pro Leu Arg Glu Thr Arg Lys Asp Pro Phe Ser Ala Ala 85 90 95 Ala Ala Glu Cys Ser Cys Arg Gln Asp Gly Leu Thr Val Ile Val Thr 100 105 110 Ala Cys Leu Thr Phe Ala Thr Gly Val Thr Val Ala Leu Val Met Gln 115 120 125 Ile Tyr Phe Gly Asp Pro Gln Ile Phe Gln Gln Gly Ala Val Val Thr 130 135 140 Asp Ala Ala Arg Cys Thr Ser Leu Gly Ile Glu Val Leu Ser Lys Gln 145 150 155 160 Gly Ser Ser Val Asp Ala Ala Val Ala Ala Ala Leu Cys Leu Gly Ile 165 170 175 Val Ala Pro His Ser Ser Gly Leu Gly Gly Gly Gly Val Met Leu Val 180 185 190 His Asp Ile Arg Arg Asn Glu Ser His Leu Ile Asp Phe Arg Glu Ser 195 200 205 Ala Pro Gly Ala Leu Arg Glu Glu Thr Leu Gln Arg Ser Trp Glu Thr 210 215 220 Lys Pro Gly Leu Leu Val Gly Val Pro Gly Met Val Lys Gly Leu His 225 230 235 240 Glu Ala His Gln Leu Tyr Gly Arg Leu Pro Trp Ser Gln Val Leu Ala 245 250 255 Phe Ala Ala Ala Val Ala Gln Asp Gly Phe Asn Val Thr His Asp Leu 260 265 270 Ala Arg Ala Leu Ala Glu Gln Leu Pro Pro Asn Met Ser Glu Arg Phe 275 280 285 Arg Glu Thr Phe Leu Pro Ser Gly Arg Pro Pro Leu Pro Gly Ser Leu 290 295 300 Leu His Arg Pro Asp Leu Ala Glu Val Leu Asp Val Leu Gly Thr Ser 305 310 315 320 Gly Pro Ala Ala Phe Tyr Ala Gly Gly Asn Leu Thr Leu Glu Met Val 325 330 335 Ala Glu Ala Gln His Ala Gly Gly Val Ile Thr Glu Glu Asp Phe Ser 340 345 350 Asn Tyr Ser Ala Leu Val Glu Lys Pro Val Cys Gly Val Tyr Arg Gly 355 360 365 His Leu Val Leu Ser Pro Pro Pro Pro His Thr Gly Pro Ala Leu Ile 370 375 380 Ser Ala Leu Asn Ile Leu Glu Gly Phe Asn Leu Thr Ser Leu Val Ser 385 390 395 400 Arg Glu Gln Ala Leu His Trp Val Ala Glu Thr Leu Lys Ile Ala Leu 405 410 415 Ala Leu Ala Ser Arg Leu Gly Asp Pro Val Tyr Asp Ser Thr Ile Thr 420 425 430 Glu Ser Met Asp Asp Met Leu Ser Lys Val Glu Ala Ala Tyr Leu Arg 435 440 445 Gly His Ile Asn Asp Ser Gln Ala Ala Pro Ala Pro Leu Leu Pro Val 450 455 460 Tyr Glu Leu Asp Gly Ala Pro Thr Ala Ala Gln Val Leu Ile Met Gly 465 470 475 480 Pro Asp Asp Phe Ile Val Ala Met Val Ser Ser Leu Asn Gln Pro Phe 485 490 495 Gly Ser Gly Leu Ile Thr Pro Ser Gly Ile Leu Leu Asn Ser Gln Met 500 505 510 Leu Asp Phe Ser Trp Pro Asn Arg Thr Ala Asn His Ser Ala Pro Ser 515 520 525 Leu Glu Asn Ser Val Gln Pro Gly Lys Arg Pro Leu Ser Phe Leu Leu 530 535 540 Pro Thr Val Val Arg Pro Ala Glu Gly Leu Cys Gly Thr Tyr Leu Ala 545 550 555 560 Leu Gly Ala Asn Gly Ala Ala Arg Gly Leu Ser Gly Leu Thr Gln Val 565 570 575 Leu Leu Asn Val Leu Thr Leu Asn Arg Asn Leu Ser Asp Ser Leu Ala 580 585 590 Arg Gly Arg Leu His Pro Asp Leu Gln Ser Asn Leu Leu Gln Val Asp 595 600 605 Ser Glu Phe Thr Glu Glu Glu Ile Glu Phe Leu Glu Ala Arg Gly His 610 615 620 His Val Glu Lys Val Asp Val Leu Ser Trp Val His Gly Ser Arg Arg 625 630 635 640 Thr Asn Asn Phe Ile Ile Ala Val Lys Asp Pro Arg Ser Pro Asp Ala 645 650 655 Ala Gly Ala Thr Ile Leu 660 12 1989 DNA Homo sapiens CDS (1)...(1989) 12 atg gcg gcg gag aac gag gcc agc cag gag agc gcc ctg ggc gcc tac 48 Met Ala Ala Glu Asn Glu Ala Ser Gln Glu Ser Ala Leu Gly Ala Tyr 1 5 10 15 tcg cca gtg gac tac atg agc atc acc agc ttc ccg cgg ctg ccc gag 96 Ser Pro Val Asp Tyr Met Ser Ile Thr Ser Phe Pro Arg Leu Pro Glu 20 25 30 gac gag ccg gcg ccc gcg gcc ccg ctg agg ggc cgc aag gac gag gac 144 Asp Glu Pro Ala Pro Ala Ala Pro Leu Arg Gly Arg Lys Asp Glu Asp 35 40 45 gcc ttt ctg gga gac ccc gac acc gac ccg gac tcc ttc ctg aag tct 192 Ala Phe Leu Gly Asp Pro Asp Thr Asp Pro Asp Ser Phe Leu Lys Ser 50 55 60 gca cgg ctg cag cgg ctg cca tcg tcg tcg tcg gag atg ggc agc caa 240 Ala Arg Leu Gln Arg Leu Pro Ser Ser Ser Ser Glu Met Gly Ser Gln 65 70 75 80 gac ggg tcg ccg cta cgc gag acg cgc aaa gac ccg ttc tcc gcc gca 288 Asp Gly Ser Pro Leu Arg Glu Thr Arg Lys Asp Pro Phe Ser Ala Ala 85 90 95 gcg gcc gag tgc tcc tgc cgc cag gat ggg ctc acg gtc atc gtc acg 336 Ala Ala Glu Cys Ser Cys Arg Gln Asp Gly Leu Thr Val Ile Val Thr 100 105 110 gcc tgt ctc acc ttc gct acc ggt gtc acc gtg gcg ctg gtc atg cag 384 Ala Cys Leu Thr Phe Ala Thr Gly Val Thr Val Ala Leu Val Met Gln 115 120 125 atc tac ttc ggg gac ccc cag atc ttc cag cag ggt gcc gtg gtg acc 432 Ile Tyr Phe Gly Asp Pro Gln Ile Phe Gln Gln Gly Ala Val Val Thr 130 135 140 gat gct gcc cgc tgc act tca ctg ggc atc gag gtg ctc agt aaa cag 480 Asp Ala Ala Arg Cys Thr Ser Leu Gly Ile Glu Val Leu Ser Lys Gln 145 150 155 160 gga tct tct gtg gac gca gcg gtg gca gca gcc ttg tgt ttg ggt atc 528 Gly Ser Ser Val Asp Ala Ala Val Ala Ala Ala Leu Cys Leu Gly Ile 165 170 175 gtg gct cca cac agt tct ggc ctg ggc ggt ggg ggc gtg atg ctg gta 576 Val Ala Pro His Ser Ser Gly Leu Gly Gly Gly Gly Val Met Leu Val 180 185 190 cat gac atc cga cga aat gag agc cac cta att gat ttc cgg gag tcc 624 His Asp Ile Arg Arg Asn Glu Ser His Leu Ile Asp Phe Arg Glu Ser 195 200 205 gca cca ggg gcc ctc agg gaa gag acc ctg caa aga tcc tgg gag acc 672 Ala Pro Gly Ala Leu Arg Glu Glu Thr Leu Gln Arg Ser Trp Glu Thr 210 215 220 aag cct ggg ctc ttg gtg ggg gtt ccc gga atg gtg aag ggg cta cat 720 Lys Pro Gly Leu Leu Val Gly Val Pro Gly Met Val Lys Gly Leu His 225 230 235 240 gaa gct cac cag ctc tat ggc agg ctg cca tgg tcc caa gtc ctg gcc 768 Glu Ala His Gln Leu Tyr Gly Arg Leu Pro Trp Ser Gln Val Leu Ala 245 250 255 ttt gca gca gct gtg gcc caa gat ggc ttc aac gtg act cat gat cta 816 Phe Ala Ala Ala Val Ala Gln Asp Gly Phe Asn Val Thr His Asp Leu 260 265 270 gcc cgt gcc ctg gct gaa cag ctg cca ccc aac atg tcc gag cgc ttc 864 Ala Arg Ala Leu Ala Glu Gln Leu Pro Pro Asn Met Ser Glu Arg Phe 275 280 285 cgg gag acg ttc ctg cca tcg ggc cgc ccg cca cta cct ggc tcg ttg 912 Arg Glu Thr Phe Leu Pro Ser Gly Arg Pro Pro Leu Pro Gly Ser Leu 290 295 300 ctg cat cgg ccc gac ctg gct gag gtg ctg gat gta ctt ggc acc tcc 960 Leu His Arg Pro Asp Leu Ala Glu Val Leu Asp Val Leu Gly Thr Ser 305 310 315 320 ggc ccg gct gcc ttc tac gca ggt ggc aac ctc aca ctg gag atg gtg 1008 Gly Pro Ala Ala Phe Tyr Ala Gly Gly Asn Leu Thr Leu Glu Met Val 325 330 335 gcc gag gct cag cac gca ggg ggt gtc ata acc gaa gag gac ttc agc 1056 Ala Glu Ala Gln His Ala Gly Gly Val Ile Thr Glu Glu Asp Phe Ser 340 345 350 aat tac agc gcc ctt gtg gag aag cct gtg tgt ggc gtg tac aga ggc 1104 Asn Tyr Ser Ala Leu Val Glu Lys Pro Val Cys Gly Val Tyr Arg Gly 355 360 365 cac ctg gtt ctt agt ccc cca cct ccg cac acg ggc cct gcc ctc atc 1152 His Leu Val Leu Ser Pro Pro Pro Pro His Thr Gly Pro Ala Leu Ile 370 375 380 agt gct ctc aac atc ctg gag ggc ttc aat ctc acc agc ctg gta tcc 1200 Ser Ala Leu Asn Ile Leu Glu Gly Phe Asn Leu Thr Ser Leu Val Ser 385 390 395 400 cga gaa cag gct ctt cac tgg gtg gca gag acc ctg aag att gca tta 1248 Arg Glu Gln Ala Leu His Trp Val Ala Glu Thr Leu Lys Ile Ala Leu 405 410 415 gcc ctg gcc agc aga ctg gga gat ccc gtc tat gat tct acc atc act 1296 Ala Leu Ala Ser Arg Leu Gly Asp Pro Val Tyr Asp Ser Thr Ile Thr 420 425 430 gag agc atg gat gac atg ctc agc aag gtg gag gcc gcc tac ctc cgg 1344 Glu Ser Met Asp Asp Met Leu Ser Lys Val Glu Ala Ala Tyr Leu Arg 435 440 445 ggc cat atc aat gac tcc cag gca gcc cct gcc cca ctc ctg cct gtc 1392 Gly His Ile Asn Asp Ser Gln Ala Ala Pro Ala Pro Leu Leu Pro Val 450 455 460 tat gaa cta gac gga gct ccc acg gct gcc cag gtg ctg atc atg gga 1440 Tyr Glu Leu Asp Gly Ala Pro Thr Ala Ala Gln Val Leu Ile Met Gly 465 470 475 480 cct gat gac ttc att gtg gcc atg gtt agc tcc ctg aac cag ccc ttt 1488 Pro Asp Asp Phe Ile Val Ala Met Val Ser Ser Leu Asn Gln Pro Phe 485 490 495 ggc agc ggc ctt atc acc ccc tcg ggg atc ctg ctc aac agc cag atg 1536 Gly Ser Gly Leu Ile Thr Pro Ser Gly Ile Leu Leu Asn Ser Gln Met 500 505 510 ctg gac ttc tcc tgg ccc aac cgg aca gct aac cac tct gca ccc agc 1584 Leu Asp Phe Ser Trp Pro Asn Arg Thr Ala Asn His Ser Ala Pro Ser 515 520 525 ctg gag aat tca gtg cag cca ggg aag cgg cca ctc tct ttc ctg ctg 1632 Leu Glu Asn Ser Val Gln Pro Gly Lys Arg Pro Leu Ser Phe Leu Leu 530 535 540 ccc aca gtg gtc cga ccc gcg gag ggg ctc tgt gga acc tac ctc gct 1680 Pro Thr Val Val Arg Pro Ala Glu Gly Leu Cys Gly Thr Tyr Leu Ala 545 550 555 560 ctg ggg gcc aat gga gct gcg cgg ggc ctc agc ggc ctg aca cag gtt 1728 Leu Gly Ala Asn Gly Ala Ala Arg Gly Leu Ser Gly Leu Thr Gln Val 565 570 575 ctg ctg aat gtc ctg acc ttg aac cgg aac ctg agt gac agc ctg gcc 1776 Leu Leu Asn Val Leu Thr Leu Asn Arg Asn Leu Ser Asp Ser Leu Ala 580 585 590 cgc ggc cgc cta cac ccg gac ctg cag tcc aac ctc ctg cag gtg gac 1824 Arg Gly Arg Leu His Pro Asp Leu Gln Ser Asn Leu Leu Gln Val Asp 595 600 605 agt gag ttc aca gag gaa gag att gag ttc ctg gaa gcc agg ggt cac 1872 Ser Glu Phe Thr Glu Glu Glu Ile Glu Phe Leu Glu Ala Arg Gly His 610 615 620 cac gtg gag aaa gta gat gtc tta tcc tgg gtc cat ggc agc cga agg 1920 His Val Glu Lys Val Asp Val Leu Ser Trp Val His Gly Ser Arg Arg 625 630 635 640 acc aac aac ttc atc atc gct gtt aag gac cct cgg agc cca gat gca 1968 Thr Asn Asn Phe Ile Ile Ala Val Lys Asp Pro Arg Ser Pro Asp Ala 645 650 655 gct gga gcc acc atc ctg tag 1989 Ala Gly Ala Thr Ile Leu * 660 13 1252 DNA Homo sapiens CDS (18)...(1187) 13 ccacgcgtcc ggaggcg atg agg gtg ttg gtg cgg cgc tgt tgg ggt cct 50 Met Arg Val Leu Val Arg Arg Cys Trp Gly Pro 1 5 10 ccg ctg gct cat ggc gcc agg cgt ggg agg ccg agt ccc cag tgg cga 98 Pro Leu Ala His Gly Ala Arg Arg Gly Arg Pro Ser Pro Gln Trp Arg 15 20 25 gca ctg gcc cga ctc ggc tgg gag gac tgc cgg gac tcc aga gtc cgc 146 Ala Leu Ala Arg Leu Gly Trp Glu Asp Cys Arg Asp Ser Arg Val Arg 30 35 40 gag aag cct ccc tgg cgg gtg ctc ttc ttc ggc acg gac cag ttc gcc 194 Glu Lys Pro Pro Trp Arg Val Leu Phe Phe Gly Thr Asp Gln Phe Ala 45 50 55 cgc gag gcg ctg cgg gcg ctg cac gcc gcc agg gaa aac aaa gaa gaa 242 Arg Glu Ala Leu Arg Ala Leu His Ala Ala Arg Glu Asn Lys Glu Glu 60 65 70 75 gag tta atc gac aaa ctg gag gtg gtc aca atg cct tcc cca tca cca 290 Glu Leu Ile Asp Lys Leu Glu Val Val Thr Met Pro Ser Pro Ser Pro 80 85 90 aaa gga ctg cca gtg aag caa tat gct gtg cag tct cag ctt ccc gta 338 Lys Gly Leu Pro Val Lys Gln Tyr Ala Val Gln Ser Gln Leu Pro Val 95 100 105 tat gag tgg ccg gat gtg gga tct gga gaa tat gat gtt gga gta gtg 386 Tyr Glu Trp Pro Asp Val Gly Ser Gly Glu Tyr Asp Val Gly Val Val 110 115 120 gct tcg ttt ggc cga ctt ttg aat gag gct ctt att ctt aaa ttt ccc 434 Ala Ser Phe Gly Arg Leu Leu Asn Glu Ala Leu Ile Leu Lys Phe Pro 125 130 135 tat ggc ata ttg aat gtt cat ccc agt tgc ctc ccg aga tgg cgt ggc 482 Tyr Gly Ile Leu Asn Val His Pro Ser Cys Leu Pro Arg Trp Arg Gly 140 145 150 155 cca gcc cct gta atc cat aca gtg ctt cac gga gac aca gtt act gga 530 Pro Ala Pro Val Ile His Thr Val Leu His Gly Asp Thr Val Thr Gly 160 165 170 gta aca att atg caa att aga cct aaa agg ttt gat gta ggc cca att 578 Val Thr Ile Met Gln Ile Arg Pro Lys Arg Phe Asp Val Gly Pro Ile 175 180 185 ctc aaa caa gaa act gtt cct gtg cca ccc aag agc act gca aag gaa 626 Leu Lys Gln Glu Thr Val Pro Val Pro Pro Lys Ser Thr Ala Lys Glu 190 195 200 ttg gaa gca gtg ttg tca aga ctg ggt gcc aac atg ctc att tca gtt 674 Leu Glu Ala Val Leu Ser Arg Leu Gly Ala Asn Met Leu Ile Ser Val 205 210 215 ttg aaa aat ttg cct gaa agt ctg agc aat gga agg cag cag cca atg 722 Leu Lys Asn Leu Pro Glu Ser Leu Ser Asn Gly Arg Gln Gln Pro Met 220 225 230 235 gag ggg gcg act tac gcc cct aag att tct gct ggt acc agt tgt ata 770 Glu Gly Ala Thr Tyr Ala Pro Lys Ile Ser Ala Gly Thr Ser Cys Ile 240 245 250 aaa tgg gag gaa caa act tca gaa caa ata ttc aga ctt tac cgt gcc 818 Lys Trp Glu Glu Gln Thr Ser Glu Gln Ile Phe Arg Leu Tyr Arg Ala 255 260 265 att gga aat ata att ccg ttg cag acg ctc tgg atg gcg aat acc att 866 Ile Gly Asn Ile Ile Pro Leu Gln Thr Leu Trp Met Ala Asn Thr Ile 270 275 280 aaa ctt ctg gat ttg gta gaa gtt aac agt tca gtc ctt gct gat cca 914 Lys Leu Leu Asp Leu Val Glu Val Asn Ser Ser Val Leu Ala Asp Pro 285 290 295 aaa tta acg gga cag gct ctt att cca gga tca gta ata tac cac aaa 962 Lys Leu Thr Gly Gln Ala Leu Ile Pro Gly Ser Val Ile Tyr His Lys 300 305 310 315 cag tca caa ata cta ttg gtt tat tgc aag gat ggt tgg att ggt gtt 1010 Gln Ser Gln Ile Leu Leu Val Tyr Cys Lys Asp Gly Trp Ile Gly Val 320 325 330 cga tca gtg atg ctc aag aaa tca cta aca gct act gac ttc tac aat 1058 Arg Ser Val Met Leu Lys Lys Ser Leu Thr Ala Thr Asp Phe Tyr Asn 335 340 345 gga tat ttg cac ccc tgg tac cag aaa aat tcc caa gct caa cca agc 1106 Gly Tyr Leu His Pro Trp Tyr Gln Lys Asn Ser Gln Ala Gln Pro Ser 350 355 360 caa tgc aga ttt cag act ctc aga ctt cca aca aag aag aag cag aaa 1154 Gln Cys Arg Phe Gln Thr Leu Arg Leu Pro Thr Lys Lys Lys Gln Lys 365 370 375 aaa act gtt gct atg caa caa tgc att gag tag ttaggaagaa gatggataaa 1207 Lys Thr Val Ala Met Gln Gln Cys Ile Glu * 380 385 aacctattac atatttgtaa tttattaaaa accttattta caagg 1252 14 389 PRT Homo sapiens 14 Met Arg Val Leu Val Arg Arg Cys Trp Gly Pro Pro Leu Ala His Gly 1 5 10 15 Ala Arg Arg Gly Arg Pro Ser Pro Gln Trp Arg Ala Leu Ala Arg Leu 20 25 30 Gly Trp Glu Asp Cys Arg Asp Ser Arg Val Arg Glu Lys Pro Pro Trp 35 40 45 Arg Val Leu Phe Phe Gly Thr Asp Gln Phe Ala Arg Glu Ala Leu Arg 50 55 60 Ala Leu His Ala Ala Arg Glu Asn Lys Glu Glu Glu Leu Ile Asp Lys 65 70 75 80 Leu Glu Val Val Thr Met Pro Ser Pro Ser Pro Lys Gly Leu Pro Val 85 90 95 Lys Gln Tyr Ala Val Gln Ser Gln Leu Pro Val Tyr Glu Trp Pro Asp 100 105 110 Val Gly Ser Gly Glu Tyr Asp Val Gly Val Val Ala Ser Phe Gly Arg 115 120 125 Leu Leu Asn Glu Ala Leu Ile Leu Lys Phe Pro Tyr Gly Ile Leu Asn 130 135 140 Val His Pro Ser Cys Leu Pro Arg Trp Arg Gly Pro Ala Pro Val Ile 145 150 155 160 His Thr Val Leu His Gly Asp Thr Val Thr Gly Val Thr Ile Met Gln 165 170 175 Ile Arg Pro Lys Arg Phe Asp Val Gly Pro Ile Leu Lys Gln Glu Thr 180 185 190 Val Pro Val Pro Pro Lys Ser Thr Ala Lys Glu Leu Glu Ala Val Leu 195 200 205 Ser Arg Leu Gly Ala Asn Met Leu Ile Ser Val Leu Lys Asn Leu Pro 210 215 220 Glu Ser Leu Ser Asn Gly Arg Gln Gln Pro Met Glu Gly Ala Thr Tyr 225 230 235 240 Ala Pro Lys Ile Ser Ala Gly Thr Ser Cys Ile Lys Trp Glu Glu Gln 245 250 255 Thr Ser Glu Gln Ile Phe Arg Leu Tyr Arg Ala Ile Gly Asn Ile Ile 260 265 270 Pro Leu Gln Thr Leu Trp Met Ala Asn Thr Ile Lys Leu Leu Asp Leu 275 280 285 Val Glu Val Asn Ser Ser Val Leu Ala Asp Pro Lys Leu Thr Gly Gln 290 295 300 Ala Leu Ile Pro Gly Ser Val Ile Tyr His Lys Gln Ser Gln Ile Leu 305 310 315 320 Leu Val Tyr Cys Lys Asp Gly Trp Ile Gly Val Arg Ser Val Met Leu 325 330 335 Lys Lys Ser Leu Thr Ala Thr Asp Phe Tyr Asn Gly Tyr Leu His Pro 340 345 350 Trp Tyr Gln Lys Asn Ser Gln Ala Gln Pro Ser Gln Cys Arg Phe Gln 355 360 365 Thr Leu Arg Leu Pro Thr Lys Lys Lys Gln Lys Lys Thr Val Ala Met 370 375 380 Gln Gln Cys Ile Glu 385 15 1170 DNA Homo sapiens CDS (1)...(1170) 15 atg agg gtg ttg gtg cgg cgc tgt tgg ggt cct ccg ctg gct cat ggc 48 Met Arg Val Leu Val Arg Arg Cys Trp Gly Pro Pro Leu Ala His Gly 1 5 10 15 gcc agg cgt ggg agg ccg agt ccc cag tgg cga gca ctg gcc cga ctc 96 Ala Arg Arg Gly Arg Pro Ser Pro Gln Trp Arg Ala Leu Ala Arg Leu 20 25 30 ggc tgg gag gac tgc cgg gac tcc aga gtc cgc gag aag cct ccc tgg 144 Gly Trp Glu Asp Cys Arg Asp Ser Arg Val Arg Glu Lys Pro Pro Trp 35 40 45 cgg gtg ctc ttc ttc ggc acg gac cag ttc gcc cgc gag gcg ctg cgg 192 Arg Val Leu Phe Phe Gly Thr Asp Gln Phe Ala Arg Glu Ala Leu Arg 50 55 60 gcg ctg cac gcc gcc agg gaa aac aaa gaa gaa gag tta atc gac aaa 240 Ala Leu His Ala Ala Arg Glu Asn Lys Glu Glu Glu Leu Ile Asp Lys 65 70 75 80 ctg gag gtg gtc aca atg cct tcc cca tca cca aaa gga ctg cca gtg 288 Leu Glu Val Val Thr Met Pro Ser Pro Ser Pro Lys Gly Leu Pro Val 85 90 95 aag caa tat gct gtg cag tct cag ctt ccc gta tat gag tgg ccg gat 336 Lys Gln Tyr Ala Val Gln Ser Gln Leu Pro Val Tyr Glu Trp Pro Asp 100 105 110 gtg gga tct gga gaa tat gat gtt gga gta gtg gct tcg ttt ggc cga 384 Val Gly Ser Gly Glu Tyr Asp Val Gly Val Val Ala Ser Phe Gly Arg 115 120 125 ctt ttg aat gag gct ctt att ctt aaa ttt ccc tat ggc ata ttg aat 432 Leu Leu Asn Glu Ala Leu Ile Leu Lys Phe Pro Tyr Gly Ile Leu Asn 130 135 140 gtt cat ccc agt tgc ctc ccg aga tgg cgt ggc cca gcc cct gta atc 480 Val His Pro Ser Cys Leu Pro Arg Trp Arg Gly Pro Ala Pro Val Ile 145 150 155 160 cat aca gtg ctt cac gga gac aca gtt act gga gta aca att atg caa 528 His Thr Val Leu His Gly Asp Thr Val Thr Gly Val Thr Ile Met Gln 165 170 175 att aga cct aaa agg ttt gat gta ggc cca att ctc aaa caa gaa act 576 Ile Arg Pro Lys Arg Phe Asp Val Gly Pro Ile Leu Lys Gln Glu Thr 180 185 190 gtt cct gtg cca ccc aag agc act gca aag gaa ttg gaa gca gtg ttg 624 Val Pro Val Pro Pro Lys Ser Thr Ala Lys Glu Leu Glu Ala Val Leu 195 200 205 tca aga ctg ggt gcc aac atg ctc att tca gtt ttg aaa aat ttg cct 672 Ser Arg Leu Gly Ala Asn Met Leu Ile Ser Val Leu Lys Asn Leu Pro 210 215 220 gaa agt ctg agc aat gga agg cag cag cca atg gag ggg gcg act tac 720 Glu Ser Leu Ser Asn Gly Arg Gln Gln Pro Met Glu Gly Ala Thr Tyr 225 230 235 240 gcc cct aag att tct gct ggt acc agt tgt ata aaa tgg gag gaa caa 768 Ala Pro Lys Ile Ser Ala Gly Thr Ser Cys Ile Lys Trp Glu Glu Gln 245 250 255 act tca gaa caa ata ttc aga ctt tac cgt gcc att gga aat ata att 816 Thr Ser Glu Gln Ile Phe Arg Leu Tyr Arg Ala Ile Gly Asn Ile Ile 260 265 270 ccg ttg cag acg ctc tgg atg gcg aat acc att aaa ctt ctg gat ttg 864 Pro Leu Gln Thr Leu Trp Met Ala Asn Thr Ile Lys Leu Leu Asp Leu 275 280 285 gta gaa gtt aac agt tca gtc ctt gct gat cca aaa tta acg gga cag 912 Val Glu Val Asn Ser Ser Val Leu Ala Asp Pro Lys Leu Thr Gly Gln 290 295 300 gct ctt att cca gga tca gta ata tac cac aaa cag tca caa ata cta 960 Ala Leu Ile Pro Gly Ser Val Ile Tyr His Lys Gln Ser Gln Ile Leu 305 310 315 320 ttg gtt tat tgc aag gat ggt tgg att ggt gtt cga tca gtg atg ctc 1008 Leu Val Tyr Cys Lys Asp Gly Trp Ile Gly Val Arg Ser Val Met Leu 325 330 335 aag aaa tca cta aca gct act gac ttc tac aat gga tat ttg cac ccc 1056 Lys Lys Ser Leu Thr Ala Thr Asp Phe Tyr Asn Gly Tyr Leu His Pro 340 345 350 tgg tac cag aaa aat tcc caa gct caa cca agc caa tgc aga ttt cag 1104 Trp Tyr Gln Lys Asn Ser Gln Ala Gln Pro Ser Gln Cys Arg Phe Gln 355 360 365 act ctc aga ctt cca aca aag aag aag cag aaa aaa act gtt gct atg 1152 Thr Leu Arg Leu Pro Thr Lys Lys Lys Gln Lys Lys Thr Val Ala Met 370 375 380 caa caa tgc att gag tag 1170 Gln Gln Cys Ile Glu * 385 16 1797 DNA Homo sapiens CDS (215)...(1189) 16 cgtccgcttt cacacgggtt gcttcggagg aatccgccgt gcaaatctgt ccgccccctt 60 ggccactgat cccccgaaga gcttctgtcg ccgctctagg aatacagaca ttgaagtttg 120 ggacaagata tttatctaac ttctgtgtca aaattagcga cctgctatgg caatgaagaa 180 agaaactgaa tttgtcattt tcacctgaag aaaa atg ata gac aaa aat caa acc 235 Met Ile Asp Lys Asn Gln Thr 1 5 tgt ggt gta gga cag gat tct gtg ccc tat atg att tgt ctg att cac 283 Cys Gly Val Gly Gln Asp Ser Val Pro Tyr Met Ile Cys Leu Ile His 10 15 20 ata ctc gaa gaa tgg ttt ggt gtg gag cag ttg gag gac tat ttg aat 331 Ile Leu Glu Glu Trp Phe Gly Val Glu Gln Leu Glu Asp Tyr Leu Asn 25 30 35 ttt gca aac tat ctc ttg tgg gtt ttt aca cca cta ata ctt tta ata 379 Phe Ala Asn Tyr Leu Leu Trp Val Phe Thr Pro Leu Ile Leu Leu Ile 40 45 50 55 ctt cct tac ttt act atc ttt ctt ctc tac ctt act att att ttc tta 427 Leu Pro Tyr Phe Thr Ile Phe Leu Leu Tyr Leu Thr Ile Ile Phe Leu 60 65 70 cac att tat aag aga aag aat gta ttg aaa gaa gcc tac tct cat aat 475 His Ile Tyr Lys Arg Lys Asn Val Leu Lys Glu Ala Tyr Ser His Asn 75 80 85 tta tgg gat ggt gca agg aaa aca gtg gca act ctg tgg gat gga cat 523 Leu Trp Asp Gly Ala Arg Lys Thr Val Ala Thr Leu Trp Asp Gly His 90 95 100 gca gcc gtt tgg cat ggt tat gaa gtt cat gga atg gaa aaa ata cca 571 Ala Ala Val Trp His Gly Tyr Glu Val His Gly Met Glu Lys Ile Pro 105 110 115 gaa gat gga cca gca ctt ata att ttt tat cat gga gct att cct ata 619 Glu Asp Gly Pro Ala Leu Ile Ile Phe Tyr His Gly Ala Ile Pro Ile 120 125 130 135 gat ttt tac tat ttc atg gct aaa ata ttt ata cac aaa ggc aga act 667 Asp Phe Tyr Tyr Phe Met Ala Lys Ile Phe Ile His Lys Gly Arg Thr 140 145 150 tgc cga gta gta gct gat cac ttt gtc ttt aaa att cca ggg ttt agt 715 Cys Arg Val Val Ala Asp His Phe Val Phe Lys Ile Pro Gly Phe Ser 155 160 165 tta tta ctg gat gtg ttt tgt gct cta cat gga cca aga gaa aaa tgt 763 Leu Leu Leu Asp Val Phe Cys Ala Leu His Gly Pro Arg Glu Lys Cys 170 175 180 gtt gaa att ctg agg agt ggc cac ttg tta gct atc tca cca ggt gga 811 Val Glu Ile Leu Arg Ser Gly His Leu Leu Ala Ile Ser Pro Gly Gly 185 190 195 gtt cga gaa gcc cta att agt gat gaa act tat aac atc gta tgg ggt 859 Val Arg Glu Ala Leu Ile Ser Asp Glu Thr Tyr Asn Ile Val Trp Gly 200 205 210 215 cat cgc aga ggc ttt gct cag gtt gca att gat gca aaa gtg ccc att 907 His Arg Arg Gly Phe Ala Gln Val Ala Ile Asp Ala Lys Val Pro Ile 220 225 230 att cct atg ttt aca caa aat att cga gaa gga ttt aga tca ctt gga 955 Ile Pro Met Phe Thr Gln Asn Ile Arg Glu Gly Phe Arg Ser Leu Gly 235 240 245 gga aca agg tta ttt agg tgg ctt tat gaa aaa ttc cgc tat cca ttt 1003 Gly Thr Arg Leu Phe Arg Trp Leu Tyr Glu Lys Phe Arg Tyr Pro Phe 250 255 260 gct cca atg tat gga ggt ttt cca gtg aag tta cgg acc tat tta ggc 1051 Ala Pro Met Tyr Gly Gly Phe Pro Val Lys Leu Arg Thr Tyr Leu Gly 265 270 275 gac ccc att ccg tat gac cca cag ata aca gcg gaa gaa tta gct gaa 1099 Asp Pro Ile Pro Tyr Asp Pro Gln Ile Thr Ala Glu Glu Leu Ala Glu 280 285 290 295 aag acg aag aat gct gtt caa gct ttg att gat aag cac caa aga ata 1147 Lys Thr Lys Asn Ala Val Gln Ala Leu Ile Asp Lys His Gln Arg Ile 300 305 310 cca gga aac att atg agt gct ttg tta gaa cgt ttt cat tga 1189 Pro Gly Asn Ile Met Ser Ala Leu Leu Glu Arg Phe His * 315 320 taacaaaggg tcaactagaa gatgatttag tacatttata ttaaatgttt gtatctaagg 1249 tactgtcttc tgaattttgt aggtcctata attagtattt tttaaaaaaa tcatgttaat 1309 aagcatcttt cacagaattc gtttctttaa aatagtcaat tttgtttttg caattgtgtc 1369 aaatactaac aaattacaca cctagtaatt cagaaaaaga tgtcttattt gtaaattcct 1429 aacaatttat gctaaacata tagattctta agtttattaa taacagcagt ttaggttaaa 1489 caaacattcc tggataatgc gttaaatttc tgtatctgtc gccctgagct gattttgaaa 1549 gatggtataa gctaggggtt agtatagttg tttaagttag aaaaaacatg ctgttgtctg 1609 cccctcattc ccttcatgac cttgggcaag tcacgtaatg tttttgtgcc tcaacaattc 1669 actttttaaa aacatgatcg tatgatgaat gatattattt tgttatttat atttactgtg 1729 attgataact gttgaaccaa aataataaaa taattaattt aaacaatgtc aaaaaaaaaa 1789 aaaaaaaa 1797 17 324 PRT Homo sapiens 17 Met Ile Asp Lys Asn Gln Thr Cys Gly Val Gly Gln Asp Ser Val Pro 1 5 10 15 Tyr Met Ile Cys Leu Ile His Ile Leu Glu Glu Trp Phe Gly Val Glu 20 25 30 Gln Leu Glu Asp Tyr Leu Asn Phe Ala Asn Tyr Leu Leu Trp Val Phe 35 40 45 Thr Pro Leu Ile Leu Leu Ile Leu Pro Tyr Phe Thr Ile Phe Leu Leu 50 55 60 Tyr Leu Thr Ile Ile Phe Leu His Ile Tyr Lys Arg Lys Asn Val Leu 65 70 75 80 Lys Glu Ala Tyr Ser His Asn Leu Trp Asp Gly Ala Arg Lys Thr Val 85 90 95 Ala Thr Leu Trp Asp Gly His Ala Ala Val Trp His Gly Tyr Glu Val 100 105 110 His Gly Met Glu Lys Ile Pro Glu Asp Gly Pro Ala Leu Ile Ile Phe 115 120 125 Tyr His Gly Ala Ile Pro Ile Asp Phe Tyr Tyr Phe Met Ala Lys Ile 130 135 140 Phe Ile His Lys Gly Arg Thr Cys Arg Val Val Ala Asp His Phe Val 145 150 155 160 Phe Lys Ile Pro Gly Phe Ser Leu Leu Leu Asp Val Phe Cys Ala Leu 165 170 175 His Gly Pro Arg Glu Lys Cys Val Glu Ile Leu Arg Ser Gly His Leu 180 185 190 Leu Ala Ile Ser Pro Gly Gly Val Arg Glu Ala Leu Ile Ser Asp Glu 195 200 205 Thr Tyr Asn Ile Val Trp Gly His Arg Arg Gly Phe Ala Gln Val Ala 210 215 220 Ile Asp Ala Lys Val Pro Ile Ile Pro Met Phe Thr Gln Asn Ile Arg 225 230 235 240 Glu Gly Phe Arg Ser Leu Gly Gly Thr Arg Leu Phe Arg Trp Leu Tyr 245 250 255 Glu Lys Phe Arg Tyr Pro Phe Ala Pro Met Tyr Gly Gly Phe Pro Val 260 265 270 Lys Leu Arg Thr Tyr Leu Gly Asp Pro Ile Pro Tyr Asp Pro Gln Ile 275 280 285 Thr Ala Glu Glu Leu Ala Glu Lys Thr Lys Asn Ala Val Gln Ala Leu 290 295 300 Ile Asp Lys His Gln Arg Ile Pro Gly Asn Ile Met Ser Ala Leu Leu 305 310 315 320 Glu Arg Phe His 18 975 DNA Homo sapiens CDS (1)...(975) 18 atg ata gac aaa aat caa acc tgt ggt gta gga cag gat tct gtg ccc 48 Met Ile Asp Lys Asn Gln Thr Cys Gly Val Gly Gln Asp Ser Val Pro 1 5 10 15 tat atg att tgt ctg att cac ata ctc gaa gaa tgg ttt ggt gtg gag 96 Tyr Met Ile Cys Leu Ile His Ile Leu Glu Glu Trp Phe Gly Val Glu 20 25 30 cag ttg gag gac tat ttg aat ttt gca aac tat ctc ttg tgg gtt ttt 144 Gln Leu Glu Asp Tyr Leu Asn Phe Ala Asn Tyr Leu Leu Trp Val Phe 35 40 45 aca cca cta ata ctt tta ata ctt cct tac ttt act atc ttt ctt ctc 192 Thr Pro Leu Ile Leu Leu Ile Leu Pro Tyr Phe Thr Ile Phe Leu Leu 50 55 60 tac ctt act att att ttc tta cac att tat aag aga aag aat gta ttg 240 Tyr Leu Thr Ile Ile Phe Leu His Ile Tyr Lys Arg Lys Asn Val Leu 65 70 75 80 aaa gaa gcc tac tct cat aat tta tgg gat ggt gca agg aaa aca gtg 288 Lys Glu Ala Tyr Ser His Asn Leu Trp Asp Gly Ala Arg Lys Thr Val 85 90 95 gca act ctg tgg gat gga cat gca gcc gtt tgg cat ggt tat gaa gtt 336 Ala Thr Leu Trp Asp Gly His Ala Ala Val Trp His Gly Tyr Glu Val 100 105 110 cat gga atg gaa aaa ata cca gaa gat gga cca gca ctt ata att ttt 384 His Gly Met Glu Lys Ile Pro Glu Asp Gly Pro Ala Leu Ile Ile Phe 115 120 125 tat cat gga gct att cct ata gat ttt tac tat ttc atg gct aaa ata 432 Tyr His Gly Ala Ile Pro Ile Asp Phe Tyr Tyr Phe Met Ala Lys Ile 130 135 140 ttt ata cac aaa ggc aga act tgc cga gta gta gct gat cac ttt gtc 480 Phe Ile His Lys Gly Arg Thr Cys Arg Val Val Ala Asp His Phe Val 145 150 155 160 ttt aaa att cca ggg ttt agt tta tta ctg gat gtg ttt tgt gct cta 528 Phe Lys Ile Pro Gly Phe Ser Leu Leu Leu Asp Val Phe Cys Ala Leu 165 170 175 cat gga cca aga gaa aaa tgt gtt gaa att ctg agg agt ggc cac ttg 576 His Gly Pro Arg Glu Lys Cys Val Glu Ile Leu Arg Ser Gly His Leu 180 185 190 tta gct atc tca cca ggt gga gtt cga gaa gcc cta att agt gat gaa 624 Leu Ala Ile Ser Pro Gly Gly Val Arg Glu Ala Leu Ile Ser Asp Glu 195 200 205 act tat aac atc gta tgg ggt cat cgc aga ggc ttt gct cag gtt gca 672 Thr Tyr Asn Ile Val Trp Gly His Arg Arg Gly Phe Ala Gln Val Ala 210 215 220 att gat gca aaa gtg ccc att att cct atg ttt aca caa aat att cga 720 Ile Asp Ala Lys Val Pro Ile Ile Pro Met Phe Thr Gln Asn Ile Arg 225 230 235 240 gaa gga ttt aga tca ctt gga gga aca agg tta ttt agg tgg ctt tat 768 Glu Gly Phe Arg Ser Leu Gly Gly Thr Arg Leu Phe Arg Trp Leu Tyr 245 250 255 gaa aaa ttc cgc tat cca ttt gct cca atg tat gga ggt ttt cca gtg 816 Glu Lys Phe Arg Tyr Pro Phe Ala Pro Met Tyr Gly Gly Phe Pro Val 260 265 270 aag tta cgg acc tat tta ggc gac ccc att ccg tat gac cca cag ata 864 Lys Leu Arg Thr Tyr Leu Gly Asp Pro Ile Pro Tyr Asp Pro Gln Ile 275 280 285 aca gcg gaa gaa tta gct gaa aag acg aag aat gct gtt caa gct ttg 912 Thr Ala Glu Glu Leu Ala Glu Lys Thr Lys Asn Ala Val Gln Ala Leu 290 295 300 att gat aag cac caa aga ata cca gga aac att atg agt gct ttg tta 960 Ile Asp Lys His Gln Arg Ile Pro Gly Asn Ile Met Ser Ala Leu Leu 305 310 315 320 gaa cgt ttt cat tga 975 Glu Arg Phe His * 19 114 PRT Artificial Sequence Amino acid consensus sequence 19 Ser Ser Ile Gly Thr Pro Val Arg Ile Arg Glu Phe Asn Asn Ser Asn 1 5 10 15 Gly Val Ser Val Ile Leu Trp Pro Cys Ser Gly Thr Thr Gly Ser Val 20 25 30 Val Trp Asp Ala Gly Val Val Leu Ser Lys Tyr Leu Leu Ser Ser Thr 35 40 45 Gln Pro His Ala Leu Ser His Ser Leu Asn Gly Lys Lys Lys Val Leu 50 55 60 Glu Leu Gly Ser Gly Thr Gly Leu Val Gly Ile Ala Ala Ala Leu Cys 65 70 75 80 Leu Gly Gly Ala Asn Val Val Leu Thr Asp Leu Pro Asp Val Leu Pro 85 90 95 Leu Leu Lys Lys Asn Val Glu Ala Asn Lys His Leu Val Gly Asn Asn 100 105 110 Ile Lys 20 102 PRT Artificial Sequence Amino acid consensus sequence 20 Ile Val Tyr Trp Asn Asn Arg Asp Gln Ile Ser Ala Leu Lys Pro Pro 1 5 10 15 Phe Asp Leu Val Ile Ala Ala Asp Val Val Tyr Ile Glu Glu Ser Val 20 25 30 Gly Gln Leu Val Thr Ala Met Glu Leu Leu Val Ala Asp Asp Gly Ala 35 40 45 Val Leu Leu Gly Tyr Gln Ile Arg Ser Pro Glu Ala Asp Lys Leu Phe 50 55 60 Trp Glu Leu Cys Asp Ile Val Phe Lys Ile Glu Lys Val Pro His Glu 65 70 75 80 His Leu His Ser Asp Tyr Ala Tyr Glu Glu Thr Asp Val Tyr Ile Phe 85 90 95 Arg Lys Lys Val Lys Lys 100 21 211 PRT Artificial Sequence Amino acid consensus sequence 21 Ser Ile Val Arg Tyr Phe Glu Leu Arg Asn Leu Ser Thr Ser Ile Pro 1 5 10 15 Leu His Glu Pro Ser Leu Thr Ala Asp Asn Leu Gly Trp Lys Thr Trp 20 25 30 Gly Ser Ser Leu Ile Leu Ser Gln Leu Val Val Asp His Leu Asp Tyr 35 40 45 Leu His Thr Thr Asn Val Asn Met Leu Ala Asn Ser Asp Ile Lys Gln 50 55 60 Ile Lys Val Leu Glu Leu Gly Ala Gly Thr Gly Leu Val Gly Leu Ser 65 70 75 80 Trp Ala Leu Lys Trp Lys Glu Leu Tyr Gly Thr Glu Asn Ile Glu Ile 85 90 95 Phe Val Thr Asp Leu Pro Glu Ile Val Thr Asn Leu Lys Lys Asn Val 100 105 110 Ser Leu Asn Asn Leu Gly Asp Phe Val Gln Ala Glu Ile Leu Asp Trp 115 120 125 Thr Asn Pro His Asp Phe Ile Asp Lys Phe Gly His Glu Asn Glu Phe 130 135 140 Asp Val Ile Leu Ile Ala Asp Pro Ile Tyr Ser Pro Gln His Pro Glu 145 150 155 160 Trp Val Val Asn Met Ile Ser Lys Phe Leu Ala Ala Ser Gly Thr Cys 165 170 175 His Leu Glu Ile Pro Leu Arg Ala Lys Tyr Ala Lys Glu Arg Glu Val 180 185 190 Leu Lys Leu Leu Leu Lys Glu Ser Asp Leu Lys Val Val Glu Glu Arg 195 200 205 His Ser Glu 210 22 191 PRT Artificial Sequence Amino acid consensus sequence 22 Asp Arg Glu Glu Ile Arg Lys Lys Leu Gly Ile Lys Glu Asp Lys Lys 1 5 10 15 Ile Ile Leu Phe Val Gly Arg Leu Val Pro Glu Lys Gly Ile Asp Leu 20 25 30 Leu Ile Glu Ala Phe Lys Lys Leu Lys Lys Lys Pro Lys Leu Leu Lys 35 40 45 Leu Asn Pro Asn Leu Lys Leu Val Ile Val Gly Gly Pro Tyr Asp Ser 50 55 60 Glu Asp Gly Glu Glu Glu Asp Glu Leu Lys Lys Leu Ala Glu Lys Leu 65 70 75 80 Gly Leu Glu Asp Asn Val Ile Phe Leu Gly Phe Val Pro Asp Glu Asp 85 90 95 Leu Pro Glu Leu Tyr Lys Ser Ala Asp Val Phe Val Leu Pro Ser Arg 100 105 110 Tyr Glu Gly Phe Gly Ile Val Leu Leu Glu Ala Met Ala Cys Gly Leu 115 120 125 Pro Val Ile Ala Thr Asn Cys Val Gly Gly Ile Pro Glu Val Val Lys 130 135 140 Asp Gly Glu Thr Gly Leu Leu Val Glu Pro Gly Gln Asp Pro Glu Ala 145 150 155 160 Leu Ala Glu Ala Ile Glu Lys Leu Leu Lys Asp Glu Glu Lys Lys Asp 165 170 175 Leu Leu Glu Leu Arg Lys Arg Leu Gly Glu Asn Ala Arg Lys Arg 180 185 190 23 49 PRT Artificial Sequence Amino acid consensus sequence 23 Val His Phe Ser Glu Ala Met Glu Lys Phe Ile His Glu Pro Ser Leu 1 5 10 15 Lys Ala Thr Met Gly Leu Ala Gly Arg Ala Arg Val Lys Glu Lys Phe 20 25 30 Ser Pro Asp Ala Phe Thr Asp Gln Leu Tyr Arg Tyr Val Thr Lys Leu 35 40 45 Leu 24 81 PRT Artificial Sequence Amino acid consensus sequence 24 Val Ala Phe Ile His Pro Asp Leu Gly Ile Gly Gly Ala Glu Arg Leu 1 5 10 15 Val Val Asp Ala Ala Val Gly Leu Gln Glu Arg Gly His Gln Val Lys 20 25 30 Ile Phe Thr Ser His His Asp Lys Ser His Cys Phe Glu Glu Thr Arg 35 40 45 Asp Gly Thr Leu Lys Val Gln Val Tyr Gly Asp Trp Leu Pro Arg Ser 50 55 60 Ile Phe Trp Gly Gly Arg Phe His Ala Ile Cys Ala Tyr Leu Arg Met 65 70 75 80 Ile 25 144 PRT Artificial Sequence Amino acid consensus sequence 25 Asn Tyr Glu Glu Lys Leu Lys Lys Leu Val Lys Glu Leu Gly Leu Glu 1 5 10 15 Asn Arg Val His Phe Leu Gly Gly Met Gly Asp Glu Glu Asp Val Ser 20 25 30 Glu Tyr Leu Lys Ser Ser Asp Ile Ile Ile Tyr Pro Ser Pro Ser Arg 35 40 45 Ser Glu Gly Phe Pro Met Val Leu Leu Glu Ala Met Ala Cys Gly Leu 50 55 60 Pro Val Ile Ala Thr Thr Thr Asp Gly Gly Gly Cys Glu Glu Ile Ile 65 70 75 80 Glu Asp Gly Glu Asn Gly Leu Leu Val Glu Pro Asn Asn Ser Asp Val 85 90 95 Glu Glu Leu Ala Glu Ala Leu Glu Lys Leu Leu Glu Asn Glu Glu Leu 100 105 110 Arg Arg Lys Met Met Gly Lys Asn Ala Arg Arg Leu Val Glu Glu Met 115 120 125 Phe Thr Ala Glu His Met Ala Lys Ala Tyr Glu Arg Phe Met Glu Lys 130 135 140 26 46 PRT Artificial Sequence Amino acid consensus sequence 26 Phe Val Ala Leu Cys Val Leu Leu Gly Trp Ser Ser Phe Asp Val Val 1 5 10 15 Leu Ala Asp Gln Val Ser Val Val Val Pro Leu Leu Lys Leu Lys Arg 20 25 30 Ser Ser Lys Val Val Phe Tyr Cys His Phe Pro Asp Leu Leu 35 40 45 27 46 PRT Artificial Sequence Amino acid consensus sequence 27 Tyr Gln Leu Ile Ile Ile Asp Gln Leu Ser Thr Cys Ile Pro Leu Leu 1 5 10 15 His Ile Phe Ser Ser Ala Thr Leu Met Phe Tyr Cys His Phe Pro Asp 20 25 30 Gln Leu Leu Ala Gln Arg Ala Gly Leu Leu Lys Lys Ile Tyr 35 40 45 28 157 PRT Artificial Sequence Amino acid consensus sequence 28 Leu Arg His Leu Val Glu Lys Gly Leu Val Ala Ala Gln Phe Leu Leu 1 5 10 15 Val Leu Gly Ala Ser Tyr Ser His Lys Asn Arg Asp Leu Ala Ile Leu 20 25 30 Ala Trp Lys Glu Leu Arg Arg Arg Gly His Asn Ile Ala Leu Val Met 35 40 45 Ala Gly Ala Val Val Ala Lys Gly Ser Ser Arg Gln Glu Glu Ala Val 50 55 60 Ala Arg Trp Gly Ala Asp Glu Glu Gln Leu Val Ile Met Pro Asp Val 65 70 75 80 Ser Ser Ala Val Arg Asn Trp Leu Leu Arg His Ala Ser Ile Val Leu 85 90 95 Tyr Pro Thr Ser Ala Glu Gly Phe Gly Leu Val Pro Phe Glu Ala Ala 100 105 110 Ser Met Gly Thr Pro Thr Ala His Val Ser Phe Gly Pro Leu Arg Glu 115 120 125 Leu Ile Asp Ser Pro Glu Leu Pro Gln Asp Trp Asp Pro Leu Arg Met 130 135 140 Ala Asp His Cys Gln Gln Leu Leu Gln Asp Pro Gln Leu 145 150 155 29 150 PRT Artificial Sequence Amino acid consensus sequence 29 Ser Ile Ile Ile Pro Thr Tyr Asn Glu Glu Lys Tyr Leu Glu Glu Cys 1 5 10 15 Leu Glu Ser Leu Leu Asn Gln Thr Thr Tyr Glu Asn Phe Glu Ile Ile 20 25 30 Val Val Asp Asp Gly Ser Thr Asp Gly Thr Val Glu Ile Leu Glu Glu 35 40 45 Tyr Ala Lys Asp Pro Arg Ile Arg Val Ile Arg Leu Glu Glu Asn Leu 50 55 60 Gly Leu Ala Ala Ala Arg Asn Ala Gly Leu Lys His Ala Thr Gly Asp 65 70 75 80 Tyr Asp Tyr Ile Ala Phe Leu Asp Ala Asp Asp Glu Val Pro Asp Trp 85 90 95 Leu Glu Lys Leu Leu Glu Leu Leu Glu Lys Asn Gly Ala Asp Ile Val 100 105 110 Ile Gly Arg Val Ile Asn Glu Asn Lys Gly Arg Leu Asn Gly Lys Leu 115 120 125 Arg Leu Leu Val Phe Leu Ile Gly Ser Asn Ala Leu Tyr Arg Arg Glu 130 135 140 Ala Leu Glu Lys Leu Leu 145 150 30 44 PRT Artificial Sequence Amino acid consensus sequence 30 Gly Thr Ile Arg Asn Leu Lys Ser Gly Leu Cys Leu Asp Val Ala Gly 1 5 10 15 Gly Ser Thr Ala Asp Gly Thr Pro Val Gln Leu Tyr Thr Cys His Gly 20 25 30 Asn Asp Gly Asn Gln Lys Trp Thr Leu Glu Lys Asp 35 40 31 44 PRT Artificial Sequence Amino acid consensus sequence 31 Gly Thr Ile Arg Asn Leu Lys Ser Gly Leu Cys Leu Asp Val Ala Gly 1 5 10 15 Gly Ser Thr Ala Asp Gly Thr Pro Val Gln Leu Tyr Thr Cys His Gly 20 25 30 Asn Asp Gly Asn Gln Lys Trp Thr Leu Glu Lys Asp 35 40 32 44 PRT Artificial Sequence Amino acid consensus sequence 32 Gly Thr Ile Arg Asn Leu Lys Ser Gly Leu Cys Leu Asp Val Ala Gly 1 5 10 15 Gly Ser Thr Ala Asp Gly Thr Pro Val Gln Leu Tyr Thr Cys His Gly 20 25 30 Asn Asp Gly Asn Gln Lys Trp Thr Leu Glu Lys Asp 35 40 33 102 PRT Artificial Sequence Amino acid consensus sequence 33 Met Gly Ser Val Thr Val Arg Tyr Phe Cys Tyr Gly Cys Leu Phe Thr 1 5 10 15 Ser Ala Thr Trp Thr Val Leu Leu Phe Val Tyr Phe Asn Phe Ser Glu 20 25 30 Val Thr Gln Pro Leu Lys Asn Val Pro Val Lys Gly Ser Gly Pro His 35 40 45 Gly Pro Ser Pro Lys Lys Phe Tyr Pro Arg Phe Thr Arg Gly Pro Ser 50 55 60 Arg Val Leu Glu Pro Gln Phe Lys Ala Asn Lys Ile Asp Asp Val Ile 65 70 75 80 Asp Ser Arg Val Glu Asp Pro Glu Glu Gly His Leu Lys Phe Ser Ser 85 90 95 Glu Leu Gly Met Ile Phe 100 34 129 PRT Artificial Sequence Amino acid consensus sequence 34 Asn Glu Glu Glu Met Lys Ala Ala Glu Glu Ser Tyr Lys Lys Tyr Ala 1 5 10 15 Phe Asn Ala Tyr Val Ser Asp Arg Ile Ser Leu Asn Arg Ser Ile Pro 20 25 30 Asp Thr Arg His Pro Glu Cys Lys Asn Lys Lys Tyr Tyr Ser Asp Asn 35 40 45 Leu Pro Thr Thr Ser Val Ile Ile Val Phe His Asn Glu Ala Trp Ser 50 55 60 Thr Leu Leu Arg Thr Val His Ser Val Ile Asn Arg Thr Pro Pro His 65 70 75 80 Leu Leu Lys Glu Ile Ile Leu Val Asp Asp Phe Ser Asp Arg Pro His 85 90 95 Leu Leu Lys Gln Lys Leu Glu Glu Tyr Val Lys Lys Lys Phe Pro Gly 100 105 110 Lys Val Lys Ile Leu Arg Asn Glu Glu Arg Glu Gly Leu Ile Arg Ala 115 120 125 Arg 35 62 PRT Artificial Sequence Amino acid consensus sequence 35 Met Asp Ile Trp Gly Gly Glu Asn Leu Glu Leu Ser Phe Arg Val Trp 1 5 10 15 Gln Cys Gly Gly Lys Leu Glu Ile Val Pro Cys Ser Arg Val Gly His 20 25 30 Ile Phe Arg Lys Gln Ser Pro Tyr Thr Phe Pro Ser Gly Ser Ser Ser 35 40 45 Asn Val Ile Ser Arg Asn Tyr Lys Arg Val Ala Glu Val Trp 50 55 60 36 41 PRT Artificial Sequence Amino acid consensus sequence 36 Asn Arg Leu Tyr Gln Val Ser Val Gly Gln Cys Leu Arg Ala Val Asp 1 5 10 15 Pro Leu Gly Gln Lys Gly Ser Val Ala Met Ala Ile Cys Asp Gly Ser 20 25 30 Ser Ser Gln Gln Trp His Leu Glu Gly 35 40 37 36 PRT Artificial Sequence Amino acid consensus sequence 37 Arg Leu Tyr Gln Val Ser Val Gly Gln Cys Leu Arg Ala Val Asp Pro 1 5 10 15 Leu Gly Gln Lys Gly Ser Val Ala Met Ala Ile Cys Asp Gly Ser Ser 20 25 30 Ser Gln Gln Trp 35 38 80 PRT Artificial Sequence Amino acid consensus sequence 38 Val Ile Asp Val Ile Asp Asp Asn Thr Phe Glu Tyr His Lys Ser Lys 1 5 10 15 Ser Ser Asp Thr Ser Arg Gly Gly Phe Asp Trp Gly Leu His Phe Lys 20 25 30 Trp His Pro Ile Pro Glu Glu Glu Arg Lys Arg Lys Lys Arg Arg Glu 35 40 45 Asp Pro Thr Glu Pro Ile Arg Ser Pro Thr Met Ala Gly Gly Leu Phe 50 55 60 Ala Ile Asp Arg Glu Tyr Phe Trp Glu Leu Gly Ser Tyr Asp Pro Gly 65 70 75 80 39 578 PRT Artificial Sequence Amino acid consensus sequence 39 Arg Arg Val Leu Lys Asp Ala His Ala Gly Gly Asn Ala Val Asp Ala 1 5 10 15 Ala Val Ala Ala Leu Phe Cys Leu Gly Val Val Glu Pro His Ala Ser 20 25 30 Gly Ile Gly Gly Gly Gly Phe Met Leu Ile Tyr Asn Leu Ala Thr Gly 35 40 45 Lys Ala Thr Val Ile Asp Phe Arg Glu Thr Ala Pro Ala Ala Ala Thr 50 55 60 Pro Asn Met Phe Leu Asp Lys Ser Gly Glu Ala Ser Lys Gln Ser Ala 65 70 75 80 Thr Gly Gly Leu Leu Ala Ile Gly Val Pro Gly Glu Val Ala Gly Leu 85 90 95 Glu Glu Ala His Lys Lys Tyr Gly Ser Thr Thr Leu Pro Trp Ala Asp 100 105 110 Leu Leu Glu Pro Ala Ile Lys Leu Ala Arg Gly Gly Phe Pro Val Ser 115 120 125 Pro Ala Leu Ala Ala Ala Leu Asp Leu Ala Glu Pro Leu Leu Leu Ser 130 135 140 Asp Ile Leu Asp Pro Gly Leu Lys Asp Ile Phe Leu Pro Asn Gly Glu 145 150 155 160 Pro Val Leu Arg Pro Gly Glu Arg Leu Val Gln Pro Asp Leu Ala Lys 165 170 175 Thr Leu Glu Leu Ile Ala Lys Glu Glu Gly Ala Asp Ala Phe Tyr Asn 180 185 190 Gly Ile Ala Ala Ser Phe Glu Leu Ala Ala Ala Leu Val Ala Asp Ile 195 200 205 Ala Lys Asn Gly Gly Ile Ile Thr Leu Glu Asp Leu Ala Asn Tyr Arg 210 215 220 Val Glu Val Arg Glu Pro Leu Ser Gly Asp Tyr Arg Gly Ala Asp Ile 225 230 235 240 Tyr Glu Val Leu Thr Met Pro Pro Pro Ser Ser Gly Gly Pro Val Leu 245 250 255 Leu Gln Ile Leu Asn Ile Leu Glu Gly Phe Asp Leu Ser Lys Tyr Ser 260 265 270 Val Gly Ser Ala Glu Tyr Lys Gly Leu Thr Val His Leu Leu Val Glu 275 280 285 Ala Met Lys Leu Ala Tyr Ala Asp Arg Asp Ala Tyr Leu Gly Asp Pro 290 295 300 Asp Phe Val Asp Val Pro Lys Val Leu Ala Lys Leu Leu Asp Lys Lys 305 310 315 320 Tyr Ala Lys Gln Arg Arg Ala Leu Ile Ser Leu Glu Lys Ala Lys Gly 325 330 335 Asp Ile Pro Ser Ser Gly Ser Leu Asp Tyr Tyr Lys Pro Gly Glu Ala 340 345 350 Ala Glu Ala Gln Asp Leu Pro Lys Glu His Gly Glu Trp Met Thr Thr 355 360 365 His Leu Ser Val Val Asp Ala Asp Gly Asn Ala Val Ser Leu Thr Ser 370 375 380 Thr Ile Asn Leu Leu Phe Gly Ser Lys Val Leu Ser Pro Gly Thr Pro 385 390 395 400 Ser Phe Gly Ile Leu Leu Asn Asn Glu Met Asp Asp Phe Ser Ser Lys 405 410 415 Leu Gly Trp Ser Pro Gly Val Gly Asn Val Phe Gly Leu Ala Pro Gly 420 425 430 Pro Ala Asn Phe Ile Glu Pro Gly Lys Arg Pro Leu Ser Ser Met Ser 435 440 445 Pro Thr Ile Val Leu Lys Lys Ser Asp Gly Lys Pro Lys Leu Val Val 450 455 460 Gly Ser Pro Gly Gly Ser Arg Ile Ile Thr Ala Val Leu Gln Thr Ile 465 470 475 480 Val Asn Val Leu Asp Tyr Gly Met Asn Leu Gln Glu Ala Val Glu Ala 485 490 495 Pro Arg Phe His His Gln Leu Leu Pro Ala Asp Arg Leu Glu Val Glu 500 505 510 Asn Phe Pro Ile Val Val Ser Glu Glu Gly Phe Ser Lys Ala Val Leu 515 520 525 Gln Glu Leu Glu Arg Arg Gly His Lys Val Glu Leu Val Pro Asp Tyr 530 535 540 Asp Lys Phe Phe Gly Ser Val Gln Ala Ile Ile Val Asp Glu Asp Gly 545 550 555 560 Glu Gly Ser Val Leu Tyr Gly Ala Ser Asp Pro Arg Arg Asn His Gly 565 570 575 Gly Glu 40 135 PRT Artificial Sequence Amino acid consensus sequence 40 Met Ala Ala Glu Asn Glu Ala Ser Gln Glu Ser Ala Leu Gly Ala Tyr 1 5 10 15 Ser Pro Val Asp Tyr Met Ser Ile Thr Ser Phe Pro Arg Leu Pro Glu 20 25 30 Asp Glu Pro Ala Pro Ala Ala Pro Leu Arg Gly Arg Lys Asp Glu Asp 35 40 45 Ala Phe Leu Gly Asp Pro Asp Thr Asp Pro Asp Ser Phe Leu Lys Ser 50 55 60 Ala Arg Leu Gln Arg Leu Pro Ser Ser Ser Ser Glu Met Gly Ser Gln 65 70 75 80 Asp Gly Ser Pro Leu Arg Glu Thr Arg Lys Asp Pro Phe Ser Ala Ala 85 90 95 Ala Ala Glu Cys Ser Cys Arg Gln Asp Gly Leu Thr Val Ile Val Thr 100 105 110 Ala Cys Leu Thr Phe Ala Thr Gly Val Thr Val Ala Leu Val Met Gln 115 120 125 Ile Tyr Phe Gly Asp Pro Gln 130 135 41 293 PRT Artificial Sequence Amino acid consensus sequence 41 Ala His Thr Ile Asp Ala Arg Glu Thr Ala Pro Ala Ala Ala Thr Glu 1 5 10 15 Asp Met Phe Glu Asn Asn Met Asp Glu Asn Asn Ser Pro Leu Lys Lys 20 25 30 Met Asp Val Thr Gly Gly Leu Ser Val Gly Val Pro Gly Glu Val Ala 35 40 45 Gly Tyr Glu Glu Ala His Lys Arg Tyr Gly Arg Leu Pro Trp Ala Gln 50 55 60 Leu Phe Gln Pro Ala Ile Lys Leu Ala Arg Glu Gly Phe Pro Val Ser 65 70 75 80 Pro Tyr Leu Ala Arg Ala Leu Glu Ser Ser Glu Glu Arg Ile Lys Leu 85 90 95 Gln Arg Pro Asp Pro Gly Trp Arg Glu Ile Phe Ala Pro Asn Gly Glu 100 105 110 Pro Leu Arg Pro Gly Glu Val Leu Lys Gln Pro Asp Leu Ala Glu Thr 115 120 125 Leu Glu Leu Ile Ala Glu Glu Gly Pro Glu Ala Phe Tyr Asn Gly Glu 130 135 140 Arg Leu Ala Glu Gln Leu Val Lys Asp Ile Gln Lys Ser Gly Gly Ile 145 150 155 160 Ile Thr Ala Glu Asp Leu Ala Asn Tyr Lys Val Lys Val Arg Glu Pro 165 170 175 Val His Ser Ser Ser Tyr Ala Arg Gly Tyr Glu Val Leu Ser Met Pro 180 185 190 Pro Pro Ser Ser Gly Gly Val Val Leu Ala Gln Val Leu Asn Ile Leu 195 200 205 Glu Gly Tyr Asn Phe Asp Met Ser Ser Val Ala Thr Pro Glu Asn Ser 210 215 220 Ala Glu Thr Tyr His Arg Leu Val Glu Ala Met Lys Phe Ala Tyr Ala 225 230 235 240 Asp Arg Ser Arg Tyr Leu Gly Asp Pro Asp Phe Val Pro Val Pro Gln 245 250 255 Asn Ala Val Glu Lys Leu Leu Ser Lys Asp Tyr Ala Lys Gln Arg Arg 260 265 270 Ala Leu Ile Pro Ser Asn Pro Gln Arg Ala Ser Pro Ser Ser Ser Leu 275 280 285 Pro Pro Gly Ala Pro 290 42 114 PRT Artificial Sequence Amino acid consensus sequence 42 Arg Pro Ala Glu Gly Leu Cys Gly Thr Tyr Leu Ala Leu Gly Ala Asn 1 5 10 15 Gly Ala Ala Arg Gly Leu Ser Gly Leu Thr Gln Val Leu Leu Asn Val 20 25 30 Leu Thr Leu Asn Arg Asn Leu Ser Asp Ser Leu Ala Arg Gly Arg Leu 35 40 45 His Pro Asp Leu Gln Ser Asn Leu Leu Gln Val Asp Ser Glu Phe Thr 50 55 60 Glu Glu Glu Ile Glu Phe Leu Glu Ala Arg Gly His His Val Glu Lys 65 70 75 80 Val Asp Val Leu Ser Trp Val His Gly Ser Arg Arg Thr Asn Asn Phe 85 90 95 Ile Ile Ala Val Lys Asp Pro Arg Ser Pro Asp Ala Ala Gly Ala Thr 100 105 110 Ile Leu 43 102 PRT Artificial Sequence Amino acid consensus sequence 43 Asp Leu Ile Val Leu Ala Gly Tyr Met Arg Ile Leu Pro Lys Glu Phe 1 5 10 15 Leu Gln Ala Phe Pro Gly Lys Ile Leu Asn Ile His Pro Ser Leu Leu 20 25 30 Pro Arg Phe Arg Gly Ala Ala Pro Ile Gln Arg Ala Leu Glu Ala Gly 35 40 45 Asp Lys Glu Thr Gly Val Thr Val His Phe Val Asp Glu Glu Leu Asp 50 55 60 Thr Gly Pro Ile Leu Ala Gln Lys Ala Val Pro Ile Leu Pro Thr Asp 65 70 75 80 Asp Thr Ser Glu Thr Leu Glu Asn Arg Val Ala Glu Leu Glu His Lys 85 90 95 Ala Leu Pro Glu Ala Leu 100 44 108 PRT Artificial Sequence Amino acid consensus sequence 44 Glu Tyr Gln Pro Asp Leu Val Val Leu Ala Gly Tyr Met Arg Ile Leu 1 5 10 15 Pro Pro Glu Phe Leu Glu Arg Tyr Pro His Gly Cys Ile Asn Ile His 20 25 30 Pro Ser Leu Leu Pro Lys Tyr Arg Gly Ala Ser Pro Ile Gln Gln Ala 35 40 45 Ile Glu Asn Gly Asp Lys Glu Thr Gly Val Thr Val His Tyr Val Asp 50 55 60 Glu Glu Glu Leu Asp Thr Gly Pro Ile Ile Ala Gln Glu Thr Val Pro 65 70 75 80 Val Glu Pro Asp Asp Thr Ala Glu Thr Thr Leu Glu Arg Val Leu Arg 85 90 95 Asp Val Glu His Glu Leu Leu Pro Glu Ala Leu Glu 100 105 45 114 PRT Artificial Sequence Amino acid consensus sequence 45 Ala Thr Tyr Ala Pro Lys Leu Lys Lys Glu Asp Gly Arg Ile Asp Trp 1 5 10 15 Asn Lys Pro Ala Glu Glu Ile His Asn Lys Ile Arg Ala Phe Ser Pro 20 25 30 Pro Trp Pro Gly Ala Trp Thr Tyr Phe Asn Gly Gln Lys Gln Lys Leu 35 40 45 Lys Ile Trp Gln Ala Lys Leu Val Asp Glu Ser Ala Ser Ser Gln Ala 50 55 60 Pro Gly Gly Thr Val Leu Ser Val Asp Lys Asn Gly Leu Leu Val Ala 65 70 75 80 Cys Gly Glu Gly Ser Val Leu Arg Leu Leu Gln Ile Gln Pro Pro Gly 85 90 95 Lys Lys Pro Met Ser Ala Lys Asp Phe Leu Asn Gly Lys Arg Glu Trp 100 105 110 Phe Lys 46 195 PRT Artificial Sequence Amino acid consensus sequence 46 Leu Glu Asn Leu Pro Lys Lys Gly Pro Ala Ile Val Val Ser Asn His 1 5 10 15 Arg Ser Tyr Leu Asp Ile Leu Val Leu Ser Ala Ala Leu Pro Arg Arg 20 25 30 Gly Pro Trp Leu Val Arg Arg Leu Val Phe Ile Ala Lys Lys Glu Leu 35 40 45 Leu Lys Val Pro Leu Leu Phe Gly Trp Leu Met Arg Leu Ala Gly Ala 50 55 60 Ile Phe Ile Asp Arg Asn Asn Arg Ala Lys Asp Ala Leu Ala Ala Ala 65 70 75 80 Asp Glu Leu Val Arg Val Leu Glu Leu Leu Arg Lys Gly Arg Ser Val 85 90 95 Leu Ile Phe Pro Glu Gly Thr Arg Ser Arg Ser Gly Glu Leu Leu Pro 100 105 110 Pro Phe Lys Lys Gly Ile Ala Ala Phe Arg Leu Ala Leu Lys Ala Gly 115 120 125 Val Pro Ile Val Pro Val Val Ile Val Ser Gly Thr Glu Glu Leu Glu 130 135 140 Pro Lys Asn Glu Ala Gly Lys Leu Leu Arg Leu Ala Arg Lys Lys Gly 145 150 155 160 Pro Val Thr Val Arg Val Leu Pro Pro Ile Pro Leu Asp Pro Glu Asp 165 170 175 Ile Lys Glu Leu Ala Glu Arg Leu Arg Asp Ile Leu Val Gln Ala Leu 180 185 190 Glu Glu Leu 195 47 216 PRT Artificial Sequence Amino acid consensus sequence 47 Gly Thr Thr Gln Arg Leu Met Pro Phe Trp Arg Trp Phe Tyr Lys Ile 1 5 10 15 Tyr His Gly Tyr Gln Val Ile Gly Leu Glu Asn Ile Pro Pro Gly Gly 20 25 30 Pro Leu Leu Val Val Tyr His His Gly Gly Ile Phe Pro Pro Pro Ile 35 40 45 Asp Met Tyr Tyr Leu Asp Trp Tyr Met Leu Leu Gly Arg Glu Arg Pro 50 55 60 Val Tyr Thr Leu Gly His Arg Phe Leu Phe Lys Gly Leu Pro Gly Trp 65 70 75 80 Gly Thr Leu Ser Glu Ala Phe His Val Ser Pro Gly Thr Val Gln Ser 85 90 95 Cys Val Ser Ala Leu Arg Asp Gly Asn Leu Val Ala Val Tyr Pro Gly 100 105 110 Gly Val Tyr Asp Ala Tyr Arg Pro Gly Asp His Tyr Tyr Glu Ile Leu 115 120 125 Trp Arg Gly Arg Lys Gly Phe Val Lys Val Ala Ile Glu Ala Gly Val 130 135 140 Pro Ile Val Pro Cys Phe Thr Gln Gly Leu Arg Glu Gly Phe Arg Gln 145 150 155 160 Val Gly Asp Cys Tyr Asp Gly Thr Trp Ile Phe Arg Thr Phe Gly Met 165 170 175 Arg Trp Tyr Asn Lys Val Asp Ile Pro Val Tyr Pro Ile Tyr Gly Gly 180 185 190 Phe Pro Trp Gly Phe Arg Thr Tyr Leu Gly Pro Pro Ile Pro Tyr Pro 195 200 205 Glu Asn Leu Thr Pro Gln Asp Leu 210 215 48 23 PRT Artificial Sequence Amino acid consensus sequence 48 Ala Ile Glu Asp Leu Ile Asn Gln His Gln Arg Leu Pro Gly Ser Ile 1 5 10 15 Leu Leu Ala Leu Leu Asp Arg 20 49 12 PRT Artificial Sequence Acyltransferase family motif 49 Asn Xaa His Arg Gln Ser Xaa Leu Tyr Ile Met Asp 1 5 10 50 9 PRT Artificial Sequence Acyltransferase family motif 50 Gly Xaa Ile Phe Phe Ile Arg Asp Arg 1 5 51 13 PRT Artificial Sequence Acyltransferase family motif 51 Phe Pro Leu Ile Glu Gly Thr Gly Arg Ser Xaa Arg Xaa 1 5 10 52 12 PRT Artificial Sequence Acyltransferase family motif 52 Val Ile Pro Xaa Ile Val Leu Ile Val Pro Val Ile 1 5 10 53 3030 DNA Homo sapiens CDS (238)...(2580) 53 ggagaattga aacccgaaca cacattgggc tcttttggca cttgactaga gctaaaacct 60 cgggattcag cgggcaagcg ttgctcagca acggcgcgta ggctgtgtgc ggttggctgg 120 agccagaccc caccccggcc tcggcccatg ctctagaggg gacgttgccc caatcctgaa 180 ggacttcggc actcgagacc tgtggatgcc gcgttgctgt ggcctgcggg ggtgatc atg 240 Met 1 aag cca ggt gct act ggc gag tcc gat ttg gcc gaa gtg ctg ccc cag 288 Lys Pro Gly Ala Thr Gly Glu Ser Asp Leu Ala Glu Val Leu Pro Gln 5 10 15 cac aag ttc gac agc aag tcc ctg gag gcc tac cta aac cag cac ttg 336 His Lys Phe Asp Ser Lys Ser Leu Glu Ala Tyr Leu Asn Gln His Leu 20 25 30 tct ggc ttt ggg gcc gaa cgt gag gct acg ctg acc att gcc cag tac 384 Ser Gly Phe Gly Ala Glu Arg Glu Ala Thr Leu Thr Ile Ala Gln Tyr 35 40 45 aga gca gga aag tcc aat cca acc ttt tat ctc cag aag ggc ttt caa 432 Arg Ala Gly Lys Ser Asn Pro Thr Phe Tyr Leu Gln Lys Gly Phe Gln 50 55 60 65 aca tat gtg ctc agg aaa aaa cca cca ggt tca ctt ctt cct aaa gca 480 Thr Tyr Val Leu Arg Lys Lys Pro Pro Gly Ser Leu Leu Pro Lys Ala 70 75 80 cat cag att gat aga gaa ttt aaa gtc cag aaa gcc ttg ttt tca att 528 His Gln Ile Asp Arg Glu Phe Lys Val Gln Lys Ala Leu Phe Ser Ile 85 90 95 gga ttc ccc gtt ccc aag cct ata ctg tac tgc agt gat act tct gtc 576 Gly Phe Pro Val Pro Lys Pro Ile Leu Tyr Cys Ser Asp Thr Ser Val 100 105 110 att gga aca gaa ttt tac gta atg gaa cat gtg cag ggt cga atc ttc 624 Ile Gly Thr Glu Phe Tyr Val Met Glu His Val Gln Gly Arg Ile Phe 115 120 125 cgt gat tta aca att cct gga ctt agc cca gca gaa cgt tca gcc ata 672 Arg Asp Leu Thr Ile Pro Gly Leu Ser Pro Ala Glu Arg Ser Ala Ile 130 135 140 145 tat gtg gcc acg gta gaa aca ttg gct cag tta cat tcc ttg aat ata 720 Tyr Val Ala Thr Val Glu Thr Leu Ala Gln Leu His Ser Leu Asn Ile 150 155 160 cag tca ctg cag ctg gaa gga tat ggt ata ggt gct ggg tac tgc aaa 768 Gln Ser Leu Gln Leu Glu Gly Tyr Gly Ile Gly Ala Gly Tyr Cys Lys 165 170 175 aga cag gta tca acc tgg aca aag caa tat caa gct gca gct cat cag 816 Arg Gln Val Ser Thr Trp Thr Lys Gln Tyr Gln Ala Ala Ala His Gln 180 185 190 gac atc cct gcc atg caa cag cta tcg gag tgg cta atg aag aac ttg 864 Asp Ile Pro Ala Met Gln Gln Leu Ser Glu Trp Leu Met Lys Asn Leu 195 200 205 ccc gat aat gac aat gaa gag aat ttg att cat gga gat ttc aga cta 912 Pro Asp Asn Asp Asn Glu Glu Asn Leu Ile His Gly Asp Phe Arg Leu 210 215 220 225 gat aac ata gtt ttc cac cct aaa gag tgt cga gtt ata gca gtg ctg 960 Asp Asn Ile Val Phe His Pro Lys Glu Cys Arg Val Ile Ala Val Leu 230 235 240 gat tgg gag ctg tca acc att ggt cat cct ttg tca gac tta gct cat 1008 Asp Trp Glu Leu Ser Thr Ile Gly His Pro Leu Ser Asp Leu Ala His 245 250 255 ttt tcc ctg ttc tac ttt tgg cca agg aca gtt cca atg ata aat caa 1056 Phe Ser Leu Phe Tyr Phe Trp Pro Arg Thr Val Pro Met Ile Asn Gln 260 265 270 ggt tct tat agt gaa aac tca ggg ata cca tca atg gaa gaa ctg att 1104 Gly Ser Tyr Ser Glu Asn Ser Gly Ile Pro Ser Met Glu Glu Leu Ile 275 280 285 tca ata tat tgc cgc tgc agg gga att aat tct att ctt cct aac tgg 1152 Ser Ile Tyr Cys Arg Cys Arg Gly Ile Asn Ser Ile Leu Pro Asn Trp 290 295 300 305 aat ttc ttt ctt gcc ctt tca tat ttt aag atg gct gga ata gca cag 1200 Asn Phe Phe Leu Ala Leu Ser Tyr Phe Lys Met Ala Gly Ile Ala Gln 310 315 320 gga gta tat agc aga tat ctt ctg gga aat aat tca tct gag gat agc 1248 Gly Val Tyr Ser Arg Tyr Leu Leu Gly Asn Asn Ser Ser Glu Asp Ser 325 330 335 ttt tta ttt gcc aat att gtg caa cct ctg gca gaa act gga cta caa 1296 Phe Leu Phe Ala Asn Ile Val Gln Pro Leu Ala Glu Thr Gly Leu Gln 340 345 350 ctc tcc aaa cga act ttc agt act gta cta cca cag att gat act act 1344 Leu Ser Lys Arg Thr Phe Ser Thr Val Leu Pro Gln Ile Asp Thr Thr 355 360 365 gga cag ttg ttt gta cag act cgg aaa ggt cag gaa gtt ctt att aag 1392 Gly Gln Leu Phe Val Gln Thr Arg Lys Gly Gln Glu Val Leu Ile Lys 370 375 380 385 gtg aag cat ttc atg aaa caa cac att ctt cca gct gaa aag gag gta 1440 Val Lys His Phe Met Lys Gln His Ile Leu Pro Ala Glu Lys Glu Val 390 395 400 act gag ttc tat gtt caa aat gaa aat tca gtg gac aag tgg gga aaa 1488 Thr Glu Phe Tyr Val Gln Asn Glu Asn Ser Val Asp Lys Trp Gly Lys 405 410 415 cct tta gtg att gat aaa ctc aag gaa atg gcc aaa gtc gag ggt ctc 1536 Pro Leu Val Ile Asp Lys Leu Lys Glu Met Ala Lys Val Glu Gly Leu 420 425 430 tgg aac ttg ttt ttg cca gct gtc agc gga ctc agc cac gtg gac tat 1584 Trp Asn Leu Phe Leu Pro Ala Val Ser Gly Leu Ser His Val Asp Tyr 435 440 445 gcc ttg att gct gaa gaa aca gga aaa tgc ttt ttt gct cca gat gtc 1632 Ala Leu Ile Ala Glu Glu Thr Gly Lys Cys Phe Phe Ala Pro Asp Val 450 455 460 465 ttt aac tgc caa gca cca gac aca ggg aat atg gag gtt ctg cac ctg 1680 Phe Asn Cys Gln Ala Pro Asp Thr Gly Asn Met Glu Val Leu His Leu 470 475 480 tat gga agt gag gaa cag aag aaa cag tgg ctt gag cct ctt ctt caa 1728 Tyr Gly Ser Glu Glu Gln Lys Lys Gln Trp Leu Glu Pro Leu Leu Gln 485 490 495 ggg aac att acc tct tgc ttc tgt atg aca gaa cct gat gta gct tca 1776 Gly Asn Ile Thr Ser Cys Phe Cys Met Thr Glu Pro Asp Val Ala Ser 500 505 510 agt gat gcc acg aat att gaa tgc agc atc caa cga gat gaa gat agc 1824 Ser Asp Ala Thr Asn Ile Glu Cys Ser Ile Gln Arg Asp Glu Asp Ser 515 520 525 tat gta att aac ggc aaa aaa tgg tgg agc agt gga gct ggg aat ccc 1872 Tyr Val Ile Asn Gly Lys Lys Trp Trp Ser Ser Gly Ala Gly Asn Pro 530 535 540 545 aag tgc aaa att gca att gtt ttg gga aga act caa aat act tct ctc 1920 Lys Cys Lys Ile Ala Ile Val Leu Gly Arg Thr Gln Asn Thr Ser Leu 550 555 560 tcc aga cac aaa cag cac agc atg att ctt gtt ccc atg aac aca cct 1968 Ser Arg His Lys Gln His Ser Met Ile Leu Val Pro Met Asn Thr Pro 565 570 575 gga gta aaa ata ata agg cct ttg tca gtt ttt ggc tac aca gat aat 2016 Gly Val Lys Ile Ile Arg Pro Leu Ser Val Phe Gly Tyr Thr Asp Asn 580 585 590 ttt cat gga gga cat ttt gag atc cat ttt aat caa gtg cga gtt cct 2064 Phe His Gly Gly His Phe Glu Ile His Phe Asn Gln Val Arg Val Pro 595 600 605 gcc aca aat cta ata cta ggt gaa ggt agg gga ttt gaa att tcc caa 2112 Ala Thr Asn Leu Ile Leu Gly Glu Gly Arg Gly Phe Glu Ile Ser Gln 610 615 620 625 ggc cgc ctt gga cct ggc aga atc cac cac tgt atg aga aca gta ggt 2160 Gly Arg Leu Gly Pro Gly Arg Ile His His Cys Met Arg Thr Val Gly 630 635 640 ttg gcg gaa cgc gct ttg cag atc atg tgt gag cgg gca aca caa agg 2208 Leu Ala Glu Arg Ala Leu Gln Ile Met Cys Glu Arg Ala Thr Gln Arg 645 650 655 ata gct ttc aag aag aag ttg tat gca cat gag gtt gtg gct cac tgg 2256 Ile Ala Phe Lys Lys Lys Leu Tyr Ala His Glu Val Val Ala His Trp 660 665 670 att gct gaa agc cgc att gcc att gag aag atc cgc ttg ttg act ctg 2304 Ile Ala Glu Ser Arg Ile Ala Ile Glu Lys Ile Arg Leu Leu Thr Leu 675 680 685 aaa gct gct cac agc atg gac act ctg ggc agt gct ggc gct aag aaa 2352 Lys Ala Ala His Ser Met Asp Thr Leu Gly Ser Ala Gly Ala Lys Lys 690 695 700 705 gag att gca atg atc aaa gtg gct gcc cca cgg gct gtc agc aaa atc 2400 Glu Ile Ala Met Ile Lys Val Ala Ala Pro Arg Ala Val Ser Lys Ile 710 715 720 gtt gac tgg gcc atc cag gtg tgc gga ggt gct ggt gtt tcc cag gat 2448 Val Asp Trp Ala Ile Gln Val Cys Gly Gly Ala Gly Val Ser Gln Asp 725 730 735 tac cct ctg gct aac atg tat gct ata acc cga gtt ttg cgt tta gca 2496 Tyr Pro Leu Ala Asn Met Tyr Ala Ile Thr Arg Val Leu Arg Leu Ala 740 745 750 gat gga cct gac gaa gtt cat ctt tca gca atc gca aca atg gag ctg 2544 Asp Gly Pro Asp Glu Val His Leu Ser Ala Ile Ala Thr Met Glu Leu 755 760 765 cgg gac caa gcc aaa aga ctg aca gcc aag ata taa ggagggtggc 2590 Arg Asp Gln Ala Lys Arg Leu Thr Ala Lys Ile * 770 775 780 actgccacat cccactggca gaaactctcc tttatacaaa cttcattggc tccaacattt 2650 gaatctcata tttttgtagc agtttgagca cagggttaat tattcatttg tggtaaagat 2710 tatagcatct attttgatca gtgggtttta ttatttcaag ggtcacacag ggttaagttc 2770 agtaagaaat gctgtagctg ttgtcattca atctagtgcc tccttgaggc caggagttca 2830 ggaccagcct gggcaacata gcgagacccc cattgctaca aaaaatttaa aaatgaaaca 2890 agtgtggtgg cacatgcttg tagtcctagc tacttgggag gctgaggcag gaggattgct 2950 tgagtctagg agtttgaggt tacagtaagc tgtgatcgtg acatggcctc cagcctgggt 3010 gaccgagtga gactgtttct 3030 54 780 PRT Homo sapiens 54 Met Lys Pro Gly Ala Thr Gly Glu Ser Asp Leu Ala Glu Val Leu Pro 1 5 10 15 Gln His Lys Phe Asp Ser Lys Ser Leu Glu Ala Tyr Leu Asn Gln His 20 25 30 Leu Ser Gly Phe Gly Ala Glu Arg Glu Ala Thr Leu Thr Ile Ala Gln 35 40 45 Tyr Arg Ala Gly Lys Ser Asn Pro Thr Phe Tyr Leu Gln Lys Gly Phe 50 55 60 Gln Thr Tyr Val Leu Arg Lys Lys Pro Pro Gly Ser Leu Leu Pro Lys 65 70 75 80 Ala His Gln Ile Asp Arg Glu Phe Lys Val Gln Lys Ala Leu Phe Ser 85 90 95 Ile Gly Phe Pro Val Pro Lys Pro Ile Leu Tyr Cys Ser Asp Thr Ser 100 105 110 Val Ile Gly Thr Glu Phe Tyr Val Met Glu His Val Gln Gly Arg Ile 115 120 125 Phe Arg Asp Leu Thr Ile Pro Gly Leu Ser Pro Ala Glu Arg Ser Ala 130 135 140 Ile Tyr Val Ala Thr Val Glu Thr Leu Ala Gln Leu His Ser Leu Asn 145 150 155 160 Ile Gln Ser Leu Gln Leu Glu Gly Tyr Gly Ile Gly Ala Gly Tyr Cys 165 170 175 Lys Arg Gln Val Ser Thr Trp Thr Lys Gln Tyr Gln Ala Ala Ala His 180 185 190 Gln Asp Ile Pro Ala Met Gln Gln Leu Ser Glu Trp Leu Met Lys Asn 195 200 205 Leu Pro Asp Asn Asp Asn Glu Glu Asn Leu Ile His Gly Asp Phe Arg 210 215 220 Leu Asp Asn Ile Val Phe His Pro Lys Glu Cys Arg Val Ile Ala Val 225 230 235 240 Leu Asp Trp Glu Leu Ser Thr Ile Gly His Pro Leu Ser Asp Leu Ala 245 250 255 His Phe Ser Leu Phe Tyr Phe Trp Pro Arg Thr Val Pro Met Ile Asn 260 265 270 Gln Gly Ser Tyr Ser Glu Asn Ser Gly Ile Pro Ser Met Glu Glu Leu 275 280 285 Ile Ser Ile Tyr Cys Arg Cys Arg Gly Ile Asn Ser Ile Leu Pro Asn 290 295 300 Trp Asn Phe Phe Leu Ala Leu Ser Tyr Phe Lys Met Ala Gly Ile Ala 305 310 315 320 Gln Gly Val Tyr Ser Arg Tyr Leu Leu Gly Asn Asn Ser Ser Glu Asp 325 330 335 Ser Phe Leu Phe Ala Asn Ile Val Gln Pro Leu Ala Glu Thr Gly Leu 340 345 350 Gln Leu Ser Lys Arg Thr Phe Ser Thr Val Leu Pro Gln Ile Asp Thr 355 360 365 Thr Gly Gln Leu Phe Val Gln Thr Arg Lys Gly Gln Glu Val Leu Ile 370 375 380 Lys Val Lys His Phe Met Lys Gln His Ile Leu Pro Ala Glu Lys Glu 385 390 395 400 Val Thr Glu Phe Tyr Val Gln Asn Glu Asn Ser Val Asp Lys Trp Gly 405 410 415 Lys Pro Leu Val Ile Asp Lys Leu Lys Glu Met Ala Lys Val Glu Gly 420 425 430 Leu Trp Asn Leu Phe Leu Pro Ala Val Ser Gly Leu Ser His Val Asp 435 440 445 Tyr Ala Leu Ile Ala Glu Glu Thr Gly Lys Cys Phe Phe Ala Pro Asp 450 455 460 Val Phe Asn Cys Gln Ala Pro Asp Thr Gly Asn Met Glu Val Leu His 465 470 475 480 Leu Tyr Gly Ser Glu Glu Gln Lys Lys Gln Trp Leu Glu Pro Leu Leu 485 490 495 Gln Gly Asn Ile Thr Ser Cys Phe Cys Met Thr Glu Pro Asp Val Ala 500 505 510 Ser Ser Asp Ala Thr Asn Ile Glu Cys Ser Ile Gln Arg Asp Glu Asp 515 520 525 Ser Tyr Val Ile Asn Gly Lys Lys Trp Trp Ser Ser Gly Ala Gly Asn 530 535 540 Pro Lys Cys Lys Ile Ala Ile Val Leu Gly Arg Thr Gln Asn Thr Ser 545 550 555 560 Leu Ser Arg His Lys Gln His Ser Met Ile Leu Val Pro Met Asn Thr 565 570 575 Pro Gly Val Lys Ile Ile Arg Pro Leu Ser Val Phe Gly Tyr Thr Asp 580 585 590 Asn Phe His Gly Gly His Phe Glu Ile His Phe Asn Gln Val Arg Val 595 600 605 Pro Ala Thr Asn Leu Ile Leu Gly Glu Gly Arg Gly Phe Glu Ile Ser 610 615 620 Gln Gly Arg Leu Gly Pro Gly Arg Ile His His Cys Met Arg Thr Val 625 630 635 640 Gly Leu Ala Glu Arg Ala Leu Gln Ile Met Cys Glu Arg Ala Thr Gln 645 650 655 Arg Ile Ala Phe Lys Lys Lys Leu Tyr Ala His Glu Val Val Ala His 660 665 670 Trp Ile Ala Glu Ser Arg Ile Ala Ile Glu Lys Ile Arg Leu Leu Thr 675 680 685 Leu Lys Ala Ala His Ser Met Asp Thr Leu Gly Ser Ala Gly Ala Lys 690 695 700 Lys Glu Ile Ala Met Ile Lys Val Ala Ala Pro Arg Ala Val Ser Lys 705 710 715 720 Ile Val Asp Trp Ala Ile Gln Val Cys Gly Gly Ala Gly Val Ser Gln 725 730 735 Asp Tyr Pro Leu Ala Asn Met Tyr Ala Ile Thr Arg Val Leu Arg Leu 740 745 750 Ala Asp Gly Pro Asp Glu Val His Leu Ser Ala Ile Ala Thr Met Glu 755 760 765 Leu Arg Asp Gln Ala Lys Arg Leu Thr Ala Lys Ile 770 775 780 55 2343 DNA Homo sapiens CDS (1)...(2343) 55 atg aag cca ggt gct act ggc gag tcc gat ttg gcc gaa gtg ctg ccc 48 Met Lys Pro Gly Ala Thr Gly Glu Ser Asp Leu Ala Glu Val Leu Pro 1 5 10 15 cag cac aag ttc gac agc aag tcc ctg gag gcc tac cta aac cag cac 96 Gln His Lys Phe Asp Ser Lys Ser Leu Glu Ala Tyr Leu Asn Gln His 20 25 30 ttg tct ggc ttt ggg gcc gaa cgt gag gct acg ctg acc att gcc cag 144 Leu Ser Gly Phe Gly Ala Glu Arg Glu Ala Thr Leu Thr Ile Ala Gln 35 40 45 tac aga gca gga aag tcc aat cca acc ttt tat ctc cag aag ggc ttt 192 Tyr Arg Ala Gly Lys Ser Asn Pro Thr Phe Tyr Leu Gln Lys Gly Phe 50 55 60 caa aca tat gtg ctc agg aaa aaa cca cca ggt tca ctt ctt cct aaa 240 Gln Thr Tyr Val Leu Arg Lys Lys Pro Pro Gly Ser Leu Leu Pro Lys 65 70 75 80 gca cat cag att gat aga gaa ttt aaa gtc cag aaa gcc ttg ttt tca 288 Ala His Gln Ile Asp Arg Glu Phe Lys Val Gln Lys Ala Leu Phe Ser 85 90 95 att gga ttc ccc gtt ccc aag cct ata ctg tac tgc agt gat act tct 336 Ile Gly Phe Pro Val Pro Lys Pro Ile Leu Tyr Cys Ser Asp Thr Ser 100 105 110 gtc att gga aca gaa ttt tac gta atg gaa cat gtg cag ggt cga atc 384 Val Ile Gly Thr Glu Phe Tyr Val Met Glu His Val Gln Gly Arg Ile 115 120 125 ttc cgt gat tta aca att cct gga ctt agc cca gca gaa cgt tca gcc 432 Phe Arg Asp Leu Thr Ile Pro Gly Leu Ser Pro Ala Glu Arg Ser Ala 130 135 140 ata tat gtg gcc acg gta gaa aca ttg gct cag tta cat tcc ttg aat 480 Ile Tyr Val Ala Thr Val Glu Thr Leu Ala Gln Leu His Ser Leu Asn 145 150 155 160 ata cag tca ctg cag ctg gaa gga tat ggt ata ggt gct ggg tac tgc 528 Ile Gln Ser Leu Gln Leu Glu Gly Tyr Gly Ile Gly Ala Gly Tyr Cys 165 170 175 aaa aga cag gta tca acc tgg aca aag caa tat caa gct gca gct cat 576 Lys Arg Gln Val Ser Thr Trp Thr Lys Gln Tyr Gln Ala Ala Ala His 180 185 190 cag gac atc cct gcc atg caa cag cta tcg gag tgg cta atg aag aac 624 Gln Asp Ile Pro Ala Met Gln Gln Leu Ser Glu Trp Leu Met Lys Asn 195 200 205 ttg ccc gat aat gac aat gaa gag aat ttg att cat gga gat ttc aga 672 Leu Pro Asp Asn Asp Asn Glu Glu Asn Leu Ile His Gly Asp Phe Arg 210 215 220 cta gat aac ata gtt ttc cac cct aaa gag tgt cga gtt ata gca gtg 720 Leu Asp Asn Ile Val Phe His Pro Lys Glu Cys Arg Val Ile Ala Val 225 230 235 240 ctg gat tgg gag ctg tca acc att ggt cat cct ttg tca gac tta gct 768 Leu Asp Trp Glu Leu Ser Thr Ile Gly His Pro Leu Ser Asp Leu Ala 245 250 255 cat ttt tcc ctg ttc tac ttt tgg cca agg aca gtt cca atg ata aat 816 His Phe Ser Leu Phe Tyr Phe Trp Pro Arg Thr Val Pro Met Ile Asn 260 265 270 caa ggt tct tat agt gaa aac tca ggg ata cca tca atg gaa gaa ctg 864 Gln Gly Ser Tyr Ser Glu Asn Ser Gly Ile Pro Ser Met Glu Glu Leu 275 280 285 att tca ata tat tgc cgc tgc agg gga att aat tct att ctt cct aac 912 Ile Ser Ile Tyr Cys Arg Cys Arg Gly Ile Asn Ser Ile Leu Pro Asn 290 295 300 tgg aat ttc ttt ctt gcc ctt tca tat ttt aag atg gct gga ata gca 960 Trp Asn Phe Phe Leu Ala Leu Ser Tyr Phe Lys Met Ala Gly Ile Ala 305 310 315 320 cag gga gta tat agc aga tat ctt ctg gga aat aat tca tct gag gat 1008 Gln Gly Val Tyr Ser Arg Tyr Leu Leu Gly Asn Asn Ser Ser Glu Asp 325 330 335 agc ttt tta ttt gcc aat att gtg caa cct ctg gca gaa act gga cta 1056 Ser Phe Leu Phe Ala Asn Ile Val Gln Pro Leu Ala Glu Thr Gly Leu 340 345 350 caa ctc tcc aaa cga act ttc agt act gta cta cca cag att gat act 1104 Gln Leu Ser Lys Arg Thr Phe Ser Thr Val Leu Pro Gln Ile Asp Thr 355 360 365 act gga cag ttg ttt gta cag act cgg aaa ggt cag gaa gtt ctt att 1152 Thr Gly Gln Leu Phe Val Gln Thr Arg Lys Gly Gln Glu Val Leu Ile 370 375 380 aag gtg aag cat ttc atg aaa caa cac att ctt cca gct gaa aag gag 1200 Lys Val Lys His Phe Met Lys Gln His Ile Leu Pro Ala Glu Lys Glu 385 390 395 400 gta act gag ttc tat gtt caa aat gaa aat tca gtg gac aag tgg gga 1248 Val Thr Glu Phe Tyr Val Gln Asn Glu Asn Ser Val Asp Lys Trp Gly 405 410 415 aaa cct tta gtg att gat aaa ctc aag gaa atg gcc aaa gtc gag ggt 1296 Lys Pro Leu Val Ile Asp Lys Leu Lys Glu Met Ala Lys Val Glu Gly 420 425 430 ctc tgg aac ttg ttt ttg cca gct gtc agc gga ctc agc cac gtg gac 1344 Leu Trp Asn Leu Phe Leu Pro Ala Val Ser Gly Leu Ser His Val Asp 435 440 445 tat gcc ttg att gct gaa gaa aca gga aaa tgc ttt ttt gct cca gat 1392 Tyr Ala Leu Ile Ala Glu Glu Thr Gly Lys Cys Phe Phe Ala Pro Asp 450 455 460 gtc ttt aac tgc caa gca cca gac aca ggg aat atg gag gtt ctg cac 1440 Val Phe Asn Cys Gln Ala Pro Asp Thr Gly Asn Met Glu Val Leu His 465 470 475 480 ctg tat gga agt gag gaa cag aag aaa cag tgg ctt gag cct ctt ctt 1488 Leu Tyr Gly Ser Glu Glu Gln Lys Lys Gln Trp Leu Glu Pro Leu Leu 485 490 495 caa ggg aac att acc tct tgc ttc tgt atg aca gaa cct gat gta gct 1536 Gln Gly Asn Ile Thr Ser Cys Phe Cys Met Thr Glu Pro Asp Val Ala 500 505 510 tca agt gat gcc acg aat att gaa tgc agc atc caa cga gat gaa gat 1584 Ser Ser Asp Ala Thr Asn Ile Glu Cys Ser Ile Gln Arg Asp Glu Asp 515 520 525 agc tat gta att aac ggc aaa aaa tgg tgg agc agt gga gct ggg aat 1632 Ser Tyr Val Ile Asn Gly Lys Lys Trp Trp Ser Ser Gly Ala Gly Asn 530 535 540 ccc aag tgc aaa att gca att gtt ttg gga aga act caa aat act tct 1680 Pro Lys Cys Lys Ile Ala Ile Val Leu Gly Arg Thr Gln Asn Thr Ser 545 550 555 560 ctc tcc aga cac aaa cag cac agc atg att ctt gtt ccc atg aac aca 1728 Leu Ser Arg His Lys Gln His Ser Met Ile Leu Val Pro Met Asn Thr 565 570 575 cct gga gta aaa ata ata agg cct ttg tca gtt ttt ggc tac aca gat 1776 Pro Gly Val Lys Ile Ile Arg Pro Leu Ser Val Phe Gly Tyr Thr Asp 580 585 590 aat ttt cat gga gga cat ttt gag atc cat ttt aat caa gtg cga gtt 1824 Asn Phe His Gly Gly His Phe Glu Ile His Phe Asn Gln Val Arg Val 595 600 605 cct gcc aca aat cta ata cta ggt gaa ggt agg gga ttt gaa att tcc 1872 Pro Ala Thr Asn Leu Ile Leu Gly Glu Gly Arg Gly Phe Glu Ile Ser 610 615 620 caa ggc cgc ctt gga cct ggc aga atc cac cac tgt atg aga aca gta 1920 Gln Gly Arg Leu Gly Pro Gly Arg Ile His His Cys Met Arg Thr Val 625 630 635 640 ggt ttg gcg gaa cgc gct ttg cag atc atg tgt gag cgg gca aca caa 1968 Gly Leu Ala Glu Arg Ala Leu Gln Ile Met Cys Glu Arg Ala Thr Gln 645 650 655 agg ata gct ttc aag aag aag ttg tat gca cat gag gtt gtg gct cac 2016 Arg Ile Ala Phe Lys Lys Lys Leu Tyr Ala His Glu Val Val Ala His 660 665 670 tgg att gct gaa agc cgc att gcc att gag aag atc cgc ttg ttg act 2064 Trp Ile Ala Glu Ser Arg Ile Ala Ile Glu Lys Ile Arg Leu Leu Thr 675 680 685 ctg aaa gct gct cac agc atg gac act ctg ggc agt gct ggc gct aag 2112 Leu Lys Ala Ala His Ser Met Asp Thr Leu Gly Ser Ala Gly Ala Lys 690 695 700 aaa gag att gca atg atc aaa gtg gct gcc cca cgg gct gtc agc aaa 2160 Lys Glu Ile Ala Met Ile Lys Val Ala Ala Pro Arg Ala Val Ser Lys 705 710 715 720 atc gtt gac tgg gcc atc cag gtg tgc gga ggt gct ggt gtt tcc cag 2208 Ile Val Asp Trp Ala Ile Gln Val Cys Gly Gly Ala Gly Val Ser Gln 725 730 735 gat tac cct ctg gct aac atg tat gct ata acc cga gtt ttg cgt tta 2256 Asp Tyr Pro Leu Ala Asn Met Tyr Ala Ile Thr Arg Val Leu Arg Leu 740 745 750 gca gat gga cct gac gaa gtt cat ctt tca gca atc gca aca atg gag 2304 Ala Asp Gly Pro Asp Glu Val His Leu Ser Ala Ile Ala Thr Met Glu 755 760 765 ctg cgg gac caa gcc aaa aga ctg aca gcc aag ata taa 2343 Leu Arg Asp Gln Ala Lys Arg Leu Thr Ala Lys Ile * 770 775 780 56 21 PRT Artificial Sequence Amino acid consensus sequence 56 Gly Ser Glu Glu Gln Lys Lys Lys Tyr Leu Pro Gln Leu Thr Ser Gly 1 5 10 15 Asp Leu Ile Gly Ala 20 57 80 PRT Artificial Sequence Amino acid consensus sequence 57 Ala Leu Thr Glu Pro Gly Ala Gly Ser Asp Val Gly Ser Leu Lys Thr 1 5 10 15 Thr Ala Glu Lys Lys Glu Gly Asp Asp Tyr Ile Leu Asn Gly Ser Lys 20 25 30 Met Trp Ile Thr Asn Gly Gly Gln Ala Asp Trp Tyr Ile Val Leu Ala 35 40 45 Val Thr Asp Pro Ala Lys Lys Val Pro Gly Lys Lys Gly Ile Thr Ala 50 55 60 Phe Leu Val Glu Lys Asp Thr Pro Gly Phe Ser Ile Gly Lys Lys Glu 65 70 75 80 58 128 PRT Artificial Sequence Amino acid consensus sequence 58 Gly Lys Gly Phe Lys Tyr Ala Met Lys Glu Leu Asp Met Glu Arg Leu 1 5 10 15 Val Ile Ala Ala Gln Ala Leu Gly Leu Ala Gln Gly Ala Leu Asp Glu 20 25 30 Ala Ile Asn Tyr Ala Lys Gln Arg Lys Gln Phe Gly Lys Pro Leu Ala 35 40 45 Asp Phe Gln Leu Ile Gln Phe Lys Leu Ala Asp Met Ala Thr Lys Leu 50 55 60 Glu Ala Ala Arg Leu Leu Val Tyr Arg Ala Ala Trp Leu Ala Asp Arg 65 70 75 80 Gly Glu Asp Ala Lys Glu Ala Leu Pro Thr Ser Lys Glu Ala Ala Met 85 90 95 Ala Lys Leu Phe Ala Ser Glu Ala Ala Met Gln Val Ala Thr Asp Ala 100 105 110 Val Gln Ile Leu Gly Gly Val Gly Tyr Thr Lys Asp Tyr Pro Val Glu 115 120 125 59 13 PRT Artificial Sequence Amino acid consensus sequence 59 Xaa Xaa Xaa Glu Xaa Xaa Xaa Gly Xaa Asp Xaa Xaa Xaa 1 5 10 60 20 PRT Artificial Sequence Amino acid consensus sequence 60 Xaa Xaa Xaa Gly Xaa Xaa Gly Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 1 5 10 15 Xaa Xaa Xaa Xaa 20 61 2004 DNA Homo sapiens CDS (119)...(1717) 61 gcactggact tgtaaacgaa aagcttcata agtccctctt tgcttagtac ttttctcgtc 60 ctttccccag ggtgcacgta accctcaagc actaggaccg tgcggaatcc aggctgcg 118 atg gca cct tca ttt acc gcc cgc att cag ttg ttc ctc ttg cgg gcg 166 Met Ala Pro Ser Phe Thr Ala Arg Ile Gln Leu Phe Leu Leu Arg Ala 1 5 10 15 cta ggc ttt ctc ata ggc tta gta ggc cga gca gct tta gtc tta ggg 214 Leu Gly Phe Leu Ile Gly Leu Val Gly Arg Ala Ala Leu Val Leu Gly 20 25 30 ggt cca aag ttt gcc tca aag acc cct cgg ccg gtg act gaa cca ttg 262 Gly Pro Lys Phe Ala Ser Lys Thr Pro Arg Pro Val Thr Glu Pro Leu 35 40 45 ctt ctg ctt tcg ggg atg cag ctg gcc aag ctg atc cga cag aga aag 310 Leu Leu Leu Ser Gly Met Gln Leu Ala Lys Leu Ile Arg Gln Arg Lys 50 55 60 gtg aaa tgt ata gat gtt gtt cag gct tat atc aac aga atc aag gac 358 Val Lys Cys Ile Asp Val Val Gln Ala Tyr Ile Asn Arg Ile Lys Asp 65 70 75 80 gtg aac cca atg atc aat gga att gtc aag tac agg ttt gag gaa gcg 406 Val Asn Pro Met Ile Asn Gly Ile Val Lys Tyr Arg Phe Glu Glu Ala 85 90 95 atg aag gag gct cat gct gta gat caa aag ctt gca gag aag cag gaa 454 Met Lys Glu Ala His Ala Val Asp Gln Lys Leu Ala Glu Lys Gln Glu 100 105 110 gat gaa gcc acc ctg gaa aat aaa tgg ccc ttc ctt ggg gtt cct ttg 502 Asp Glu Ala Thr Leu Glu Asn Lys Trp Pro Phe Leu Gly Val Pro Leu 115 120 125 aca gtc aag gaa gct ttc cag cta caa gga atg ccc aat tct tct gga 550 Thr Val Lys Glu Ala Phe Gln Leu Gln Gly Met Pro Asn Ser Ser Gly 130 135 140 ctc atg aac cgt cgt gat gcc att gcc aaa aca gat gcc act gtg gtg 598 Leu Met Asn Arg Arg Asp Ala Ile Ala Lys Thr Asp Ala Thr Val Val 145 150 155 160 gca tta ctg aag gga gct ggt gcc att cct ctt ggc ata acc aac tgt 646 Ala Leu Leu Lys Gly Ala Gly Ala Ile Pro Leu Gly Ile Thr Asn Cys 165 170 175 agt gag ttg tgt atg tgg tat gaa tcc agt aac aag atc tat ggc cga 694 Ser Glu Leu Cys Met Trp Tyr Glu Ser Ser Asn Lys Ile Tyr Gly Arg 180 185 190 tca aac aac cca tat gat tta cag cat att gta ggt gga agt tct ggt 742 Ser Asn Asn Pro Tyr Asp Leu Gln His Ile Val Gly Gly Ser Ser Gly 195 200 205 ggt gag ggc tgc aca ctg gca gct gcc tgc tca gtt att ggt gtg ggc 790 Gly Glu Gly Cys Thr Leu Ala Ala Ala Cys Ser Val Ile Gly Val Gly 210 215 220 tct gat att ggt ggt agc att cga atg cct gct ttc ttc aat ggt ata 838 Ser Asp Ile Gly Gly Ser Ile Arg Met Pro Ala Phe Phe Asn Gly Ile 225 230 235 240 ttt gga cac aag cct tct cca ggt gtg gtt ccc aac aaa ggt cag ttt 886 Phe Gly His Lys Pro Ser Pro Gly Val Val Pro Asn Lys Gly Gln Phe 245 250 255 ccc ttg gct gtg gga gcc cag gag ttg ttt ctg tgc act ggt cct atg 934 Pro Leu Ala Val Gly Ala Gln Glu Leu Phe Leu Cys Thr Gly Pro Met 260 265 270 tgc cgc tat gct gaa gac ctg gcc ccc atg ttg aag gtc atg gca gga 982 Cys Arg Tyr Ala Glu Asp Leu Ala Pro Met Leu Lys Val Met Ala Gly 275 280 285 cct ggg atc aaa agg tta aaa cta gac aca aag gta cat tta aaa gac 1030 Pro Gly Ile Lys Arg Leu Lys Leu Asp Thr Lys Val His Leu Lys Asp 290 295 300 tta aaa ttt tac tgg atg gaa cat gat gga ggc tca ttt tta atg tcc 1078 Leu Lys Phe Tyr Trp Met Glu His Asp Gly Gly Ser Phe Leu Met Ser 305 310 315 320 aaa gtg gac caa gat ctc att atg act cag aaa aag gtt gtg gtt cac 1126 Lys Val Asp Gln Asp Leu Ile Met Thr Gln Lys Lys Val Val Val His 325 330 335 ctt gaa act att cta gga gcc tca gtt caa cat gtt aaa ctg aag aaa 1174 Leu Glu Thr Ile Leu Gly Ala Ser Val Gln His Val Lys Leu Lys Lys 340 345 350 atg aag tac tct ttt cag ttg tgg atc gca atg atg tca gca aag gga 1222 Met Lys Tyr Ser Phe Gln Leu Trp Ile Ala Met Met Ser Ala Lys Gly 355 360 365 cat gat ggg aag gaa cct gtg aaa ttt gta gat ttg ctt ggt gac cat 1270 His Asp Gly Lys Glu Pro Val Lys Phe Val Asp Leu Leu Gly Asp His 370 375 380 ggg aaa cat gtc agt cct ctg tgg gag ttg atc aaa tgg tgc ctg ggt 1318 Gly Lys His Val Ser Pro Leu Trp Glu Leu Ile Lys Trp Cys Leu Gly 385 390 395 400 ctg tca gtg tac acc atc cct tcc att gga ctg gct ttg ttg gaa gaa 1366 Leu Ser Val Tyr Thr Ile Pro Ser Ile Gly Leu Ala Leu Leu Glu Glu 405 410 415 aag ctc aga tat agc aat gag aaa tac caa aag ttt aag gca gtg gaa 1414 Lys Leu Arg Tyr Ser Asn Glu Lys Tyr Gln Lys Phe Lys Ala Val Glu 420 425 430 gaa agc ctg cgt aaa gag ctg gtg gat atg cta ggt gat gat ggt gtg 1462 Glu Ser Leu Arg Lys Glu Leu Val Asp Met Leu Gly Asp Asp Gly Val 435 440 445 ttc tta tat ccc tca cat ccc aca gtg gca cct aag cat cat gtc cct 1510 Phe Leu Tyr Pro Ser His Pro Thr Val Ala Pro Lys His His Val Pro 450 455 460 cta aca cgg ccc ttc aac ttt gct tac aca ggt gtc ttc agt gcc ctg 1558 Leu Thr Arg Pro Phe Asn Phe Ala Tyr Thr Gly Val Phe Ser Ala Leu 465 470 475 480 ggt ttg cct gtg acc caa tgc cca ctg gga ctg aat gcc aaa gga ctc 1606 Gly Leu Pro Val Thr Gln Cys Pro Leu Gly Leu Asn Ala Lys Gly Leu 485 490 495 cct tta ggc atc cag gtt gtg gct gga ccc ttt aat gat cat ctg acc 1654 Pro Leu Gly Ile Gln Val Val Ala Gly Pro Phe Asn Asp His Leu Thr 500 505 510 ctg gct gtg gcc cag tac ttg gag aaa act ttt ggg ggc tgg gtc tgt 1702 Leu Ala Val Ala Gln Tyr Leu Glu Lys Thr Phe Gly Gly Trp Val Cys 515 520 525 cca gga aag ttt tag gaggaccttc tgcaaggtta atgtgtgtgt gtgtttgtgt 1757 Pro Gly Lys Phe * 530 tcgtgtggtg gtgtttctat taattgggtg aaaccaagca ccagcagaca agcagagaaa 1817 caactgggga atttattgac tcatttagtt attctttcta cttttatttc cttctctaac 1877 tgttggtctt actaaaatgg taatatttgc ttcttgcttt tatgttactg gaaaattagg 1937 acatgtaaat ggataagtgc aataaagttt cctaaatgct gaaaaaaaaa aaaaaaaaaa 1997 aggccgc 2004 62 532 PRT Homo sapiens 62 Met Ala Pro Ser Phe Thr Ala Arg Ile Gln Leu Phe Leu Leu Arg Ala 1 5 10 15 Leu Gly Phe Leu Ile Gly Leu Val Gly Arg Ala Ala Leu Val Leu Gly 20 25 30 Gly Pro Lys Phe Ala Ser Lys Thr Pro Arg Pro Val Thr Glu Pro Leu 35 40 45 Leu Leu Leu Ser Gly Met Gln Leu Ala Lys Leu Ile Arg Gln Arg Lys 50 55 60 Val Lys Cys Ile Asp Val Val Gln Ala Tyr Ile Asn Arg Ile Lys Asp 65 70 75 80 Val Asn Pro Met Ile Asn Gly Ile Val Lys Tyr Arg Phe Glu Glu Ala 85 90 95 Met Lys Glu Ala His Ala Val Asp Gln Lys Leu Ala Glu Lys Gln Glu 100 105 110 Asp Glu Ala Thr Leu Glu Asn Lys Trp Pro Phe Leu Gly Val Pro Leu 115 120 125 Thr Val Lys Glu Ala Phe Gln Leu Gln Gly Met Pro Asn Ser Ser Gly 130 135 140 Leu Met Asn Arg Arg Asp Ala Ile Ala Lys Thr Asp Ala Thr Val Val 145 150 155 160 Ala Leu Leu Lys Gly Ala Gly Ala Ile Pro Leu Gly Ile Thr Asn Cys 165 170 175 Ser Glu Leu Cys Met Trp Tyr Glu Ser Ser Asn Lys Ile Tyr Gly Arg 180 185 190 Ser Asn Asn Pro Tyr Asp Leu Gln His Ile Val Gly Gly Ser Ser Gly 195 200 205 Gly Glu Gly Cys Thr Leu Ala Ala Ala Cys Ser Val Ile Gly Val Gly 210 215 220 Ser Asp Ile Gly Gly Ser Ile Arg Met Pro Ala Phe Phe Asn Gly Ile 225 230 235 240 Phe Gly His Lys Pro Ser Pro Gly Val Val Pro Asn Lys Gly Gln Phe 245 250 255 Pro Leu Ala Val Gly Ala Gln Glu Leu Phe Leu Cys Thr Gly Pro Met 260 265 270 Cys Arg Tyr Ala Glu Asp Leu Ala Pro Met Leu Lys Val Met Ala Gly 275 280 285 Pro Gly Ile Lys Arg Leu Lys Leu Asp Thr Lys Val His Leu Lys Asp 290 295 300 Leu Lys Phe Tyr Trp Met Glu His Asp Gly Gly Ser Phe Leu Met Ser 305 310 315 320 Lys Val Asp Gln Asp Leu Ile Met Thr Gln Lys Lys Val Val Val His 325 330 335 Leu Glu Thr Ile Leu Gly Ala Ser Val Gln His Val Lys Leu Lys Lys 340 345 350 Met Lys Tyr Ser Phe Gln Leu Trp Ile Ala Met Met Ser Ala Lys Gly 355 360 365 His Asp Gly Lys Glu Pro Val Lys Phe Val Asp Leu Leu Gly Asp His 370 375 380 Gly Lys His Val Ser Pro Leu Trp Glu Leu Ile Lys Trp Cys Leu Gly 385 390 395 400 Leu Ser Val Tyr Thr Ile Pro Ser Ile Gly Leu Ala Leu Leu Glu Glu 405 410 415 Lys Leu Arg Tyr Ser Asn Glu Lys Tyr Gln Lys Phe Lys Ala Val Glu 420 425 430 Glu Ser Leu Arg Lys Glu Leu Val Asp Met Leu Gly Asp Asp Gly Val 435 440 445 Phe Leu Tyr Pro Ser His Pro Thr Val Ala Pro Lys His His Val Pro 450 455 460 Leu Thr Arg Pro Phe Asn Phe Ala Tyr Thr Gly Val Phe Ser Ala Leu 465 470 475 480 Gly Leu Pro Val Thr Gln Cys Pro Leu Gly Leu Asn Ala Lys Gly Leu 485 490 495 Pro Leu Gly Ile Gln Val Val Ala Gly Pro Phe Asn Asp His Leu Thr 500 505 510 Leu Ala Val Ala Gln Tyr Leu Glu Lys Thr Phe Gly Gly Trp Val Cys 515 520 525 Pro Gly Lys Phe 530 63 1599 DNA Homo sapiens CDS (1)...(1599) 63 atg gca cct tca ttt acc gcc cgc att cag ttg ttc ctc ttg cgg gcg 48 Met Ala Pro Ser Phe Thr Ala Arg Ile Gln Leu Phe Leu Leu Arg Ala 1 5 10 15 cta ggc ttt ctc ata ggc tta gta ggc cga gca gct tta gtc tta ggg 96 Leu Gly Phe Leu Ile Gly Leu Val Gly Arg Ala Ala Leu Val Leu Gly 20 25 30 ggt cca aag ttt gcc tca aag acc cct cgg ccg gtg act gaa cca ttg 144 Gly Pro Lys Phe Ala Ser Lys Thr Pro Arg Pro Val Thr Glu Pro Leu 35 40 45 ctt ctg ctt tcg ggg atg cag ctg gcc aag ctg atc cga cag aga aag 192 Leu Leu Leu Ser Gly Met Gln Leu Ala Lys Leu Ile Arg Gln Arg Lys 50 55 60 gtg aaa tgt ata gat gtt gtt cag gct tat atc aac aga atc aag gac 240 Val Lys Cys Ile Asp Val Val Gln Ala Tyr Ile Asn Arg Ile Lys Asp 65 70 75 80 gtg aac cca atg atc aat gga att gtc aag tac agg ttt gag gaa gcg 288 Val Asn Pro Met Ile Asn Gly Ile Val Lys Tyr Arg Phe Glu Glu Ala 85 90 95 atg aag gag gct cat gct gta gat caa aag ctt gca gag aag cag gaa 336 Met Lys Glu Ala His Ala Val Asp Gln Lys Leu Ala Glu Lys Gln Glu 100 105 110 gat gaa gcc acc ctg gaa aat aaa tgg ccc ttc ctt ggg gtt cct ttg 384 Asp Glu Ala Thr Leu Glu Asn Lys Trp Pro Phe Leu Gly Val Pro Leu 115 120 125 aca gtc aag gaa gct ttc cag cta caa gga atg ccc aat tct tct gga 432 Thr Val Lys Glu Ala Phe Gln Leu Gln Gly Met Pro Asn Ser Ser Gly 130 135 140 ctc atg aac cgt cgt gat gcc att gcc aaa aca gat gcc act gtg gtg 480 Leu Met Asn Arg Arg Asp Ala Ile Ala Lys Thr Asp Ala Thr Val Val 145 150 155 160 gca tta ctg aag gga gct ggt gcc att cct ctt ggc ata acc aac tgt 528 Ala Leu Leu Lys Gly Ala Gly Ala Ile Pro Leu Gly Ile Thr Asn Cys 165 170 175 agt gag ttg tgt atg tgg tat gaa tcc agt aac aag atc tat ggc cga 576 Ser Glu Leu Cys Met Trp Tyr Glu Ser Ser Asn Lys Ile Tyr Gly Arg 180 185 190 tca aac aac cca tat gat tta cag cat att gta ggt gga agt tct ggt 624 Ser Asn Asn Pro Tyr Asp Leu Gln His Ile Val Gly Gly Ser Ser Gly 195 200 205 ggt gag ggc tgc aca ctg gca gct gcc tgc tca gtt att ggt gtg ggc 672 Gly Glu Gly Cys Thr Leu Ala Ala Ala Cys Ser Val Ile Gly Val Gly 210 215 220 tct gat att ggt ggt agc att cga atg cct gct ttc ttc aat ggt ata 720 Ser Asp Ile Gly Gly Ser Ile Arg Met Pro Ala Phe Phe Asn Gly Ile 225 230 235 240 ttt gga cac aag cct tct cca ggt gtg gtt ccc aac aaa ggt cag ttt 768 Phe Gly His Lys Pro Ser Pro Gly Val Val Pro Asn Lys Gly Gln Phe 245 250 255 ccc ttg gct gtg gga gcc cag gag ttg ttt ctg tgc act ggt cct atg 816 Pro Leu Ala Val Gly Ala Gln Glu Leu Phe Leu Cys Thr Gly Pro Met 260 265 270 tgc cgc tat gct gaa gac ctg gcc ccc atg ttg aag gtc atg gca gga 864 Cys Arg Tyr Ala Glu Asp Leu Ala Pro Met Leu Lys Val Met Ala Gly 275 280 285 cct ggg atc aaa agg tta aaa cta gac aca aag gta cat tta aaa gac 912 Pro Gly Ile Lys Arg Leu Lys Leu Asp Thr Lys Val His Leu Lys Asp 290 295 300 tta aaa ttt tac tgg atg gaa cat gat gga ggc tca ttt tta atg tcc 960 Leu Lys Phe Tyr Trp Met Glu His Asp Gly Gly Ser Phe Leu Met Ser 305 310 315 320 aaa gtg gac caa gat ctc att atg act cag aaa aag gtt gtg gtt cac 1008 Lys Val Asp Gln Asp Leu Ile Met Thr Gln Lys Lys Val Val Val His 325 330 335 ctt gaa act att cta gga gcc tca gtt caa cat gtt aaa ctg aag aaa 1056 Leu Glu Thr Ile Leu Gly Ala Ser Val Gln His Val Lys Leu Lys Lys 340 345 350 atg aag tac tct ttt cag ttg tgg atc gca atg atg tca gca aag gga 1104 Met Lys Tyr Ser Phe Gln Leu Trp Ile Ala Met Met Ser Ala Lys Gly 355 360 365 cat gat ggg aag gaa cct gtg aaa ttt gta gat ttg ctt ggt gac cat 1152 His Asp Gly Lys Glu Pro Val Lys Phe Val Asp Leu Leu Gly Asp His 370 375 380 ggg aaa cat gtc agt cct ctg tgg gag ttg atc aaa tgg tgc ctg ggt 1200 Gly Lys His Val Ser Pro Leu Trp Glu Leu Ile Lys Trp Cys Leu Gly 385 390 395 400 ctg tca gtg tac acc atc cct tcc att gga ctg gct ttg ttg gaa gaa 1248 Leu Ser Val Tyr Thr Ile Pro Ser Ile Gly Leu Ala Leu Leu Glu Glu 405 410 415 aag ctc aga tat agc aat gag aaa tac caa aag ttt aag gca gtg gaa 1296 Lys Leu Arg Tyr Ser Asn Glu Lys Tyr Gln Lys Phe Lys Ala Val Glu 420 425 430 gaa agc ctg cgt aaa gag ctg gtg gat atg cta ggt gat gat ggt gtg 1344 Glu Ser Leu Arg Lys Glu Leu Val Asp Met Leu Gly Asp Asp Gly Val 435 440 445 ttc tta tat ccc tca cat ccc aca gtg gca cct aag cat cat gtc cct 1392 Phe Leu Tyr Pro Ser His Pro Thr Val Ala Pro Lys His His Val Pro 450 455 460 cta aca cgg ccc ttc aac ttt gct tac aca ggt gtc ttc agt gcc ctg 1440 Leu Thr Arg Pro Phe Asn Phe Ala Tyr Thr Gly Val Phe Ser Ala Leu 465 470 475 480 ggt ttg cct gtg acc caa tgc cca ctg gga ctg aat gcc aaa gga ctc 1488 Gly Leu Pro Val Thr Gln Cys Pro Leu Gly Leu Asn Ala Lys Gly Leu 485 490 495 cct tta ggc atc cag gtt gtg gct gga ccc ttt aat gat cat ctg acc 1536 Pro Leu Gly Ile Gln Val Val Ala Gly Pro Phe Asn Asp His Leu Thr 500 505 510 ctg gct gtg gcc cag tac ttg gag aaa act ttt ggg ggc tgg gtc tgt 1584 Leu Ala Val Ala Gln Tyr Leu Glu Lys Thr Phe Gly Gly Trp Val Cys 515 520 525 cca gga aag ttt tag 1599 Pro Gly Lys Phe * 530 64 218 PRT Artificial Sequence Amino acid consensus sequence 64 Glu Leu Val Glu Ala Phe Leu Ala Arg Ile Glu Ala Ala Asn Pro Lys 1 5 10 15 Leu Asn Val Thr Ala Phe Val Thr Val Phe Phe Glu Glu Ala Leu Ala 20 25 30 Ala Ala Lys Ala Ala Asp Lys Arg Arg Ala Arg Lys Arg Gly Gly Glu 35 40 45 Lys Gly Pro Leu His Gly Val Pro Ile Ala Leu Lys Asp Asn Ile Asp 50 55 60 Val Lys Gly Val Pro Thr Thr Ala Gly Ser Lys Ala Leu Glu Gly Tyr 65 70 75 80 Pro Pro Pro Tyr Asp Ala Thr Val Val Glu Arg Leu Arg Ala Ala Gly 85 90 95 Ala Val Ile Leu Gly Lys Thr Asn Met Asp Glu Phe Ala Met Gly Ser 100 105 110 Thr Thr Glu Asn Ser Ala Phe Gly Pro Thr Arg Asn Pro Trp Asp Leu 115 120 125 Ser Arg Thr Pro Gly Gly Ser Ser Gly Gly Ser Ala Ala Ala Val Ala 130 135 140 Ala Gly Leu Val Pro Leu Ala Ile Gly Thr Asp Thr Gly Gly Ser Ile 145 150 155 160 Arg Ile Pro Ala Ala Phe Cys Gly Leu Val Gly Leu Lys Pro Thr Tyr 165 170 175 Gly Arg Val Ser Arg Tyr Gly Val Val Gly Ser Val Glu Pro Leu Ser 180 185 190 Ser Ser Leu Asp Gln Val Gly Pro Leu Ala Arg Ser Val Glu Asp Ala 195 200 205 Ala Leu Leu Leu Asp Val Ile Ala Gly Tyr 210 215 65 127 PRT Artificial Sequence Amino acid consensus sequence 65 Gly Tyr Ser Asp Ala Tyr Glu Tyr Leu Lys Ala Gln Lys Val Arg Arg 1 5 10 15 Leu Leu Arg Arg Glu Phe Asp Gly Leu Phe Glu Glu His Gly Val Asp 20 25 30 Val Leu Ile Ser Pro Thr Thr Pro Thr Pro Ala Pro Arg Ile Gly Glu 35 40 45 Pro Asp Lys Leu Ile Ser Glu Ala Asp Asp Tyr Thr Val Leu Tyr Leu 50 55 60 Leu Asp Asp Phe Thr Ala Asn Thr Val Pro Ala Asn Leu Ala Gly Leu 65 70 75 80 Pro Ala Ile Ser Val Pro Val Gly Phe Ser Pro Glu Asp Ser Trp Asp 85 90 95 Ala Leu Val Lys Glu Tyr Leu Pro Glu Gly Tyr Val Gly Leu Pro Val 100 105 110 Gly Leu Gln Ile Ile Gly Lys Pro Gly Asp Glu Glu Thr Leu Leu 115 120 125 66 22 PRT Artificial Sequence Amidase signature motif 66 Gly Xaa Ser Xaa Xaa Gly Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 1 5 10 15 Ser Xaa Arg Xaa Pro Xaa 20 67 1816 DNA Homo sapiens CDS (84)...(1436) 67 cacgcgtccg ggggccttgg aggcccagcc cgcgcggcga cgtctccgcg tggcgtcacg 60 gcaccgactg acggccaccc acc atg gcc gca gac cag cgc ccg aag gcc gac 113 Met Ala Ala Asp Gln Arg Pro Lys Ala Asp 1 5 10 acg ctg gcc ctg agg caa cgg ctc atc agc tct tcc tgc aga ccc ttt 161 Thr Leu Ala Leu Arg Gln Arg Leu Ile Ser Ser Ser Cys Arg Pro Phe 15 20 25 ttt ccc gag gat cct gtt aag att gtc cgg gcc caa ggg cag tac atg 209 Phe Pro Glu Asp Pro Val Lys Ile Val Arg Ala Gln Gly Gln Tyr Met 30 35 40 tac gat gaa cag ggg gca gaa tac atc gat tgc atc agc aat gtg gcg 257 Tyr Asp Glu Gln Gly Ala Glu Tyr Ile Asp Cys Ile Ser Asn Val Ala 45 50 55 cac gtt ggg cac tgc cac cct ctc gtg gtc caa gca gca cat gag cag 305 His Val Gly His Cys His Pro Leu Val Val Gln Ala Ala His Glu Gln 60 65 70 aac cag gtg ctc aac acc aac agc cgg tac ctg cat gac aac atc gtg 353 Asn Gln Val Leu Asn Thr Asn Ser Arg Tyr Leu His Asp Asn Ile Val 75 80 85 90 gac tat gcg cag agg ctg tca gag acc ctg ccg gag cag ctc tgt gtg 401 Asp Tyr Ala Gln Arg Leu Ser Glu Thr Leu Pro Glu Gln Leu Cys Val 95 100 105 ttc tat ttc ctg aat tct ggg tca gaa gcc aat gac ctg gcc ctg agg 449 Phe Tyr Phe Leu Asn Ser Gly Ser Glu Ala Asn Asp Leu Ala Leu Arg 110 115 120 ctg gct cgc cac tac acg gga cac cag gac gtg gtg gta tta gat cat 497 Leu Ala Arg His Tyr Thr Gly His Gln Asp Val Val Val Leu Asp His 125 130 135 gcg tat cac ggc cac ctg agc tcc ctg att gac atc agt ccc tac aag 545 Ala Tyr His Gly His Leu Ser Ser Leu Ile Asp Ile Ser Pro Tyr Lys 140 145 150 ttc cgc aac ctg gat ggc cag aag gag tgg gtc cac gtg gca cct ctc 593 Phe Arg Asn Leu Asp Gly Gln Lys Glu Trp Val His Val Ala Pro Leu 155 160 165 170 cca gac acc tac cgg ggc ccc tac cgg gag gac cac ccc aac cca gct 641 Pro Asp Thr Tyr Arg Gly Pro Tyr Arg Glu Asp His Pro Asn Pro Ala 175 180 185 atg gcc tat gcc aac gag gtg aaa cgt gtg gtc agc agt gca cag gag 689 Met Ala Tyr Ala Asn Glu Val Lys Arg Val Val Ser Ser Ala Gln Glu 190 195 200 aag ggc agg aag att gca gcc ttc ttc gct gag tct ctg ccc agt gtg 737 Lys Gly Arg Lys Ile Ala Ala Phe Phe Ala Glu Ser Leu Pro Ser Val 205 210 215 gga ggg cag atc att ccc cct gct ggc tac ttc tcc caa gtg gca gag 785 Gly Gly Gln Ile Ile Pro Pro Ala Gly Tyr Phe Ser Gln Val Ala Glu 220 225 230 cac atc cgc aag gcc gga ggg gtc ttt gtt gca gat gag atc cag gtt 833 His Ile Arg Lys Ala Gly Gly Val Phe Val Ala Asp Glu Ile Gln Val 235 240 245 250 ggc ttt ggc cgg gta ggc aag cac ttc tgg gcc ttc cag ctc cag gga 881 Gly Phe Gly Arg Val Gly Lys His Phe Trp Ala Phe Gln Leu Gln Gly 255 260 265 aaa gac ttc gtc cct gac atc gtc acc atg ggc aag tcc att ggc aac 929 Lys Asp Phe Val Pro Asp Ile Val Thr Met Gly Lys Ser Ile Gly Asn 270 275 280 ggc cac cct gtt gcc tgc gtg gcc gca acc cag cct gtg gcg agg gca 977 Gly His Pro Val Ala Cys Val Ala Ala Thr Gln Pro Val Ala Arg Ala 285 290 295 ttt gaa gcc acc ggc gtt gag tac ttc aac acg ttt ggg ggc agc cca 1025 Phe Glu Ala Thr Gly Val Glu Tyr Phe Asn Thr Phe Gly Gly Ser Pro 300 305 310 gtg tcc tgc gct gtg ggg ctg gcc gtc ctg aat gtc ttg gag aag gag 1073 Val Ser Cys Ala Val Gly Leu Ala Val Leu Asn Val Leu Glu Lys Glu 315 320 325 330 cag ctc cag gat cat gcc acc agt gta ggc agc ttc ctg atg cag ctc 1121 Gln Leu Gln Asp His Ala Thr Ser Val Gly Ser Phe Leu Met Gln Leu 335 340 345 ctc ggg cag caa aaa atc aaa cat ccc atc gtc ggg gat gtc agg ggt 1169 Leu Gly Gln Gln Lys Ile Lys His Pro Ile Val Gly Asp Val Arg Gly 350 355 360 gtt ggg ctc ttc att ggt gtg gat ctg atc aaa gat gag gcc aca agg 1217 Val Gly Leu Phe Ile Gly Val Asp Leu Ile Lys Asp Glu Ala Thr Arg 365 370 375 aca cca gca act gaa gag gct gcc tac ttg gta tca agg ctg aag gag 1265 Thr Pro Ala Thr Glu Glu Ala Ala Tyr Leu Val Ser Arg Leu Lys Glu 380 385 390 aac tac gtt ttg ctg agc act gat ggc cct ggg agg aac atc ctg aag 1313 Asn Tyr Val Leu Leu Ser Thr Asp Gly Pro Gly Arg Asn Ile Leu Lys 395 400 405 410 ttt aag ccc cca atg tgc ttc agc ctg gac aat gca cgg cag gtg gtg 1361 Phe Lys Pro Pro Met Cys Phe Ser Leu Asp Asn Ala Arg Gln Val Val 415 420 425 gca aag ctg gat gcc att ctg act gac atg gaa gag aag gtg aga agt 1409 Ala Lys Leu Asp Ala Ile Leu Thr Asp Met Glu Glu Lys Val Arg Ser 430 435 440 tgt gaa acg ctg agg ctc cag ccc taa gccagccctg ctctgcctaa 1456 Cys Glu Thr Leu Arg Leu Gln Pro * 445 450 gtgtactcca gaagaaactc atctcatcca aatacacgct attgagaagg cgagcctgac 1516 ctccctctta cagataaagt cagctttcag aggctcaggg tgggggggcc tgcccgaggc 1576 cataatgcta cccaccccct cctcctaacc actggtctgt tggaataacc cagatgtctg 1636 catcccctca agtcagtcaa tttcctttct gtccactggg ggtggaatgg ggtagggtgg 1696 gatactttaa agtgctcctg cttaaataaa ttagaccaga ccagtgtatt tctaaagaaa 1756 atcctgacat gcacacccat taaaaatagt acattttaca gtgaaaaaaa aaaaaaaagg 1816 68 450 PRT Homo sapiens 68 Met Ala Ala Asp Gln Arg Pro Lys Ala Asp Thr Leu Ala Leu Arg Gln 1 5 10 15 Arg Leu Ile Ser Ser Ser Cys Arg Pro Phe Phe Pro Glu Asp Pro Val 20 25 30 Lys Ile Val Arg Ala Gln Gly Gln Tyr Met Tyr Asp Glu Gln Gly Ala 35 40 45 Glu Tyr Ile Asp Cys Ile Ser Asn Val Ala His Val Gly His Cys His 50 55 60 Pro Leu Val Val Gln Ala Ala His Glu Gln Asn Gln Val Leu Asn Thr 65 70 75 80 Asn Ser Arg Tyr Leu His Asp Asn Ile Val Asp Tyr Ala Gln Arg Leu 85 90 95 Ser Glu Thr Leu Pro Glu Gln Leu Cys Val Phe Tyr Phe Leu Asn Ser 100 105 110 Gly Ser Glu Ala Asn Asp Leu Ala Leu Arg Leu Ala Arg His Tyr Thr 115 120 125 Gly His Gln Asp Val Val Val Leu Asp His Ala Tyr His Gly His Leu 130 135 140 Ser Ser Leu Ile Asp Ile Ser Pro Tyr Lys Phe Arg Asn Leu Asp Gly 145 150 155 160 Gln Lys Glu Trp Val His Val Ala Pro Leu Pro Asp Thr Tyr Arg Gly 165 170 175 Pro Tyr Arg Glu Asp His Pro Asn Pro Ala Met Ala Tyr Ala Asn Glu 180 185 190 Val Lys Arg Val Val Ser Ser Ala Gln Glu Lys Gly Arg Lys Ile Ala 195 200 205 Ala Phe Phe Ala Glu Ser Leu Pro Ser Val Gly Gly Gln Ile Ile Pro 210 215 220 Pro Ala Gly Tyr Phe Ser Gln Val Ala Glu His Ile Arg Lys Ala Gly 225 230 235 240 Gly Val Phe Val Ala Asp Glu Ile Gln Val Gly Phe Gly Arg Val Gly 245 250 255 Lys His Phe Trp Ala Phe Gln Leu Gln Gly Lys Asp Phe Val Pro Asp 260 265 270 Ile Val Thr Met Gly Lys Ser Ile Gly Asn Gly His Pro Val Ala Cys 275 280 285 Val Ala Ala Thr Gln Pro Val Ala Arg Ala Phe Glu Ala Thr Gly Val 290 295 300 Glu Tyr Phe Asn Thr Phe Gly Gly Ser Pro Val Ser Cys Ala Val Gly 305 310 315 320 Leu Ala Val Leu Asn Val Leu Glu Lys Glu Gln Leu Gln Asp His Ala 325 330 335 Thr Ser Val Gly Ser Phe Leu Met Gln Leu Leu Gly Gln Gln Lys Ile 340 345 350 Lys His Pro Ile Val Gly Asp Val Arg Gly Val Gly Leu Phe Ile Gly 355 360 365 Val Asp Leu Ile Lys Asp Glu Ala Thr Arg Thr Pro Ala Thr Glu Glu 370 375 380 Ala Ala Tyr Leu Val Ser Arg Leu Lys Glu Asn Tyr Val Leu Leu Ser 385 390 395 400 Thr Asp Gly Pro Gly Arg Asn Ile Leu Lys Phe Lys Pro Pro Met Cys 405 410 415 Phe Ser Leu Asp Asn Ala Arg Gln Val Val Ala Lys Leu Asp Ala Ile 420 425 430 Leu Thr Asp Met Glu Glu Lys Val Arg Ser Cys Glu Thr Leu Arg Leu 435 440 445 Gln Pro 450 69 1353 DNA Homo sapiens CDS (1)...(1353) 69 atg gcc gca gac cag cgc ccg aag gcc gac acg ctg gcc ctg agg caa 48 Met Ala Ala Asp Gln Arg Pro Lys Ala Asp Thr Leu Ala Leu Arg Gln 1 5 10 15 cgg ctc atc agc tct tcc tgc aga ccc ttt ttt ccc gag gat cct gtt 96 Arg Leu Ile Ser Ser Ser Cys Arg Pro Phe Phe Pro Glu Asp Pro Val 20 25 30 aag att gtc cgg gcc caa ggg cag tac atg tac gat gaa cag ggg gca 144 Lys Ile Val Arg Ala Gln Gly Gln Tyr Met Tyr Asp Glu Gln Gly Ala 35 40 45 gaa tac atc gat tgc atc agc aat gtg gcg cac gtt ggg cac tgc cac 192 Glu Tyr Ile Asp Cys Ile Ser Asn Val Ala His Val Gly His Cys His 50 55 60 cct ctc gtg gtc caa gca gca cat gag cag aac cag gtg ctc aac acc 240 Pro Leu Val Val Gln Ala Ala His Glu Gln Asn Gln Val Leu Asn Thr 65 70 75 80 aac agc cgg tac ctg cat gac aac atc gtg gac tat gcg cag agg ctg 288 Asn Ser Arg Tyr Leu His Asp Asn Ile Val Asp Tyr Ala Gln Arg Leu 85 90 95 tca gag acc ctg ccg gag cag ctc tgt gtg ttc tat ttc ctg aat tct 336 Ser Glu Thr Leu Pro Glu Gln Leu Cys Val Phe Tyr Phe Leu Asn Ser 100 105 110 ggg tca gaa gcc aat gac ctg gcc ctg agg ctg gct cgc cac tac acg 384 Gly Ser Glu Ala Asn Asp Leu Ala Leu Arg Leu Ala Arg His Tyr Thr 115 120 125 gga cac cag gac gtg gtg gta tta gat cat gcg tat cac ggc cac ctg 432 Gly His Gln Asp Val Val Val Leu Asp His Ala Tyr His Gly His Leu 130 135 140 agc tcc ctg att gac atc agt ccc tac aag ttc cgc aac ctg gat ggc 480 Ser Ser Leu Ile Asp Ile Ser Pro Tyr Lys Phe Arg Asn Leu Asp Gly 145 150 155 160 cag aag gag tgg gtc cac gtg gca cct ctc cca gac acc tac cgg ggc 528 Gln Lys Glu Trp Val His Val Ala Pro Leu Pro Asp Thr Tyr Arg Gly 165 170 175 ccc tac cgg gag gac cac ccc aac cca gct atg gcc tat gcc aac gag 576 Pro Tyr Arg Glu Asp His Pro Asn Pro Ala Met Ala Tyr Ala Asn Glu 180 185 190 gtg aaa cgt gtg gtc agc agt gca cag gag aag ggc agg aag att gca 624 Val Lys Arg Val Val Ser Ser Ala Gln Glu Lys Gly Arg Lys Ile Ala 195 200 205 gcc ttc ttc gct gag tct ctg ccc agt gtg gga ggg cag atc att ccc 672 Ala Phe Phe Ala Glu Ser Leu Pro Ser Val Gly Gly Gln Ile Ile Pro 210 215 220 cct gct ggc tac ttc tcc caa gtg gca gag cac atc cgc aag gcc gga 720 Pro Ala Gly Tyr Phe Ser Gln Val Ala Glu His Ile Arg Lys Ala Gly 225 230 235 240 ggg gtc ttt gtt gca gat gag atc cag gtt ggc ttt ggc cgg gta ggc 768 Gly Val Phe Val Ala Asp Glu Ile Gln Val Gly Phe Gly Arg Val Gly 245 250 255 aag cac ttc tgg gcc ttc cag ctc cag gga aaa gac ttc gtc cct gac 816 Lys His Phe Trp Ala Phe Gln Leu Gln Gly Lys Asp Phe Val Pro Asp 260 265 270 atc gtc acc atg ggc aag tcc att ggc aac ggc cac cct gtt gcc tgc 864 Ile Val Thr Met Gly Lys Ser Ile Gly Asn Gly His Pro Val Ala Cys 275 280 285 gtg gcc gca acc cag cct gtg gcg agg gca ttt gaa gcc acc ggc gtt 912 Val Ala Ala Thr Gln Pro Val Ala Arg Ala Phe Glu Ala Thr Gly Val 290 295 300 gag tac ttc aac acg ttt ggg ggc agc cca gtg tcc tgc gct gtg ggg 960 Glu Tyr Phe Asn Thr Phe Gly Gly Ser Pro Val Ser Cys Ala Val Gly 305 310 315 320 ctg gcc gtc ctg aat gtc ttg gag aag gag cag ctc cag gat cat gcc 1008 Leu Ala Val Leu Asn Val Leu Glu Lys Glu Gln Leu Gln Asp His Ala 325 330 335 acc agt gta ggc agc ttc ctg atg cag ctc ctc ggg cag caa aaa atc 1056 Thr Ser Val Gly Ser Phe Leu Met Gln Leu Leu Gly Gln Gln Lys Ile 340 345 350 aaa cat ccc atc gtc ggg gat gtc agg ggt gtt ggg ctc ttc att ggt 1104 Lys His Pro Ile Val Gly Asp Val Arg Gly Val Gly Leu Phe Ile Gly 355 360 365 gtg gat ctg atc aaa gat gag gcc aca agg aca cca gca act gaa gag 1152 Val Asp Leu Ile Lys Asp Glu Ala Thr Arg Thr Pro Ala Thr Glu Glu 370 375 380 gct gcc tac ttg gta tca agg ctg aag gag aac tac gtt ttg ctg agc 1200 Ala Ala Tyr Leu Val Ser Arg Leu Lys Glu Asn Tyr Val Leu Leu Ser 385 390 395 400 act gat ggc cct ggg agg aac atc ctg aag ttt aag ccc cca atg tgc 1248 Thr Asp Gly Pro Gly Arg Asn Ile Leu Lys Phe Lys Pro Pro Met Cys 405 410 415 ttc agc ctg gac aat gca cgg cag gtg gtg gca aag ctg gat gcc att 1296 Phe Ser Leu Asp Asn Ala Arg Gln Val Val Ala Lys Leu Asp Ala Ile 420 425 430 ctg act gac atg gaa gag aag gtg aga agt tgt gaa acg ctg agg ctc 1344 Leu Thr Asp Met Glu Glu Lys Val Arg Ser Cys Glu Thr Leu Arg Leu 435 440 445 cag ccc taa 1353 Gln Pro * 450 70 483 PRT Artificial Sequence Amino acid consensus sequence 70 Ser Val Ala Arg Gly Asn Tyr Gly Pro Leu Pro Val Leu Ile Thr Arg 1 5 10 15 Ala Lys Gly Val Trp Leu Thr Asp Val Asp Gly Arg Glu Tyr Leu Asp 20 25 30 Phe Leu Ser Gly Ile Ala Val Ala Asn Leu Gly His Cys His Pro Lys 35 40 45 Val Val Gln Ala Val Lys Glu Gln Ala Asp Lys Leu Thr His Thr Ser 50 55 60 Arg Ala Phe Leu Thr His Glu Pro Ala Leu Asp Phe Val Glu Lys Leu 65 70 75 80 Ala Glu Lys Leu Ala Ser Leu Thr Pro Gly Asp Gly Leu Asp Arg Val 85 90 95 Phe Phe Met Asn Ser Gly Ser Glu Ala Asn Glu Thr Ala Leu Lys Leu 100 105 110 Ala Arg Ala Tyr Ala Arg Gln Lys Gly Lys Val Pro Glu Lys Phe Ser 115 120 125 Glu Glu Leu Glu Ser Met Leu Asn Gln Pro Gly Thr Gly Lys Thr Lys 130 135 140 Ile Ile Ala Phe Ser Gly Ala Phe His Gly Arg Thr Leu Gly Ala Leu 145 150 155 160 Ser Val Thr Gly Ser Lys Lys Gly Tyr Arg Lys Leu Phe Gly Pro Leu 165 170 175 Leu Pro Gly Val Val Tyr Ala Ala Ala Asp Thr Leu Phe Ala Pro Tyr 180 185 190 Asn Asp Pro Ser Leu Tyr Arg Pro Pro Phe Glu Glu Gly Lys Glu Asn 195 200 205 Ala Ser Glu Gly Leu Glu Ala Lys Leu Glu Glu Ala Leu Glu Asp Leu 210 215 220 Ile Glu Glu Tyr Lys Lys Lys Asp Asp Glu Ile Ala Ala Val Ile Val 225 230 235 240 Glu Pro Ile Val Gln Gly Glu Gly Gly Val Ile Pro Pro Pro Pro Gly 245 250 255 Phe Leu Ala Gly Leu Arg Glu Leu Cys Lys Lys His Gly Val Leu Leu 260 265 270 Ile Ala Asp Glu Val Gln Thr Gly Phe Gly Arg Thr Gly Lys Leu Phe 275 280 285 Ala Cys Glu His Xaa Xaa Xaa Asp Gly Val Thr Pro Pro Asp Ile Met 290 295 300 Thr Leu Ala Lys Ala Leu Gly Gly Gly Val Leu Pro Leu Ala Ala Val 305 310 315 320 Ile Gly Arg Ala Glu Ile Met Gln Ala Phe Phe Asp Ala Pro Gly Gly 325 330 335 Glu Ala Lys Pro Phe Leu His Gly Thr Thr Phe Gly Gly Asn Pro Leu 340 345 350 Ala Cys Ala Ala Ala Leu Ala Thr Leu Lys Val Leu Glu Glu Glu Asn 355 360 365 Leu Leu Gln Asn Ala Gln Glu Lys Gly Asp Tyr Leu Arg Lys Gly Leu 370 375 380 Leu Glu Leu Ala Lys Lys Tyr Pro Asp Val Ile Gly Asp Val Arg Gly 385 390 395 400 Lys Gly Leu Met Ile Gly Ile Glu Ile Val Glu Asp Arg Val Thr Lys 405 410 415 Glu Pro Ala Ala Lys Pro Ser Asp Glu Glu Leu Val Ala Asp Ile Ile 420 425 430 Lys Ala Ala Leu Glu Lys Gly Leu Leu Ile Leu Pro Ala Gly Tyr Val 435 440 445 Arg Asn Gly Gly Asn Val Ile Arg Phe Ala Pro Pro Leu Thr Ile Thr 450 455 460 Asp Glu Glu Ile Asp Glu Gly Leu Asp Ala Leu Lys Lys Ala Leu Ala 465 470 475 480 Lys Ala Leu 71 166 PRT Artificial Sequence Amino acid consensus sequence 71 Tyr Leu His His Glu Ile His Asp Tyr Ala Glu Arg Leu Thr Ala Lys 1 5 10 15 Met Pro Gly Pro Leu Lys Val Val Phe Phe Val Asn Ser Gly Ser Glu 20 25 30 Ala Asn Asp Leu Ala Met Met Met Ala Arg Asn Tyr Thr Gly His Gln 35 40 45 Asp Val Ile Ser Leu Arg Asn Ala Tyr His Gly Met Ser Pro Thr Thr 50 55 60 Met Gly Leu Thr Asn Leu Gly Thr Trp Lys Tyr Pro Xaa Leu Pro Gly 65 70 75 80 Val Gln Ser Gly Ile His His Val Met Asn Pro Asp Pro Tyr Arg Gly 85 90 95 Ile Trp Gly Ser Asp Gly Glu Lys Xaa Xaa Xaa Xaa Tyr Ala Lys Asp 100 105 110 Val Gln Xaa Thr Phe Lys Tyr Tyr Gly Pro Arg Gly Xaa Lys Val Ala 115 120 125 Ala Phe Ile Ala Glu Ser Ile Gln Gly Val Gly Gly Thr Val Gln Leu 130 135 140 Pro Pro Gly Tyr Leu Lys Ala Val Tyr Asp Ile Val Arg Ser Ala Gly 145 150 155 160 Gly Val Cys Ile Ala Asp 165 72 57 PRT Artificial Sequence Amino acid consensus sequence 72 Ser Thr Tyr Gly Gly Asn Pro Leu Ala Cys Ala Ala Ala Leu Ala Thr 1 5 10 15 Leu Glu Ile Ile Glu Glu Glu Asn Leu Val Glu Arg Ala Gln Glu Leu 20 25 30 Gly Glu Tyr Leu Arg Glu Arg Leu Leu Glu Met Gln Glu Glu His His 35 40 45 Pro Ile Val Gly Asp Val Arg Thr Val 50 55 73 1971 DNA Homo sapiens misc_feature (1)...(1971) n = A,T,C or G 73 nnnnnnnnnn nnnnnnnnnn nnaccaggac cgctcggcgn nnnnnnnnnn nnnnnnnnnn 60 nnnnnnnnnn nnnnnnnnnc nncatggccg cggacacggc gccnnaaggc cgtcactctg 120 gacctgagac gtcggctgct cagctcttcc tgcagactct tttttcctga ggatcctgtt 180 aagattatcc gaggccaagg gcagtacctg tacgatgagc aagggcgaga gtacctggac 240 tgtatcaaca acgtggctca tgttgggcac tgccacccta ccgtggtcca agccgcacat 300 gaacagaacc tagtgctcaa caccaacagc cgctacctgc atggcaacat cgtggactat 360 gcccagaggc tgtcggagac cctgccggag cagctctctg tgttttactt cttgaattct 420 gggtcagaag ccaacgacct ggccttgaga ctagctcgac agtacacggg acaccaggat 480 gtggtggtat tagaccatgc ttatcatggt cacctgagct ccctgatcga catcagtccc 540 tacaagttcc ggaatctggg tggccagaag gaatgggtcc atgtggctcc tctcccagac 600 acctaccggg gcccttacag ggaggaccac cccaacccag cagaggccta tgccaacgag 660 gtgaagcacg tcatcagcag tgcacagcag aagggcagga agatcgcagc cttcttcgct 720 gagtctctgc ccagtgtgag tggacagatc attcctcctg ctggctactt ctcccaggtg 780 gctgagcaca tccacagagc tccgcaaggc cggagggctc tttgtggcag atgagatcca 840 ggttggtttt ggccgcatag gcaagcactt ttgggccttc cagctggagg gagaagactt 900 tgttcccgac attgtcacca tgggcaagtc catcggcaat ggtcaccctg ttgcctgcat 960 ggccactacc caagctgtgt caagggcatt tgaagctacc ggtgtagaat acttcaacac 1020 gtttggtggc aaccccgtat cctgtgctgt ggggctagca gtcctagatg tcttgaaaac 1080 agaacagctc caggctcacg ccactaatgt ccaccagtgt gggcagtttc cttctggagc 1140 acctcaccca gcagaaagcc aagcacccta tcattggaga tgtcaggggc actggactct 1200 tcatcggtgt ggatctcatc aaagatgaga ccctgaggac accagcaact gaagaggcgg 1260 aatatttggt ctccaggcta aaggaaaact acattttact gagcattgat ggccctggaa 1320 agaatattct gaagttcaag cccccaatgt gcttcaacgt tgacaatgca caacatgtgg 1380 tagcaaagct ggatgacatt ctaacagaca tggaagaaaa agtaagaagt tgtgagaccc 1440 tgaggatcaa gcnnnacnnn nnncccnnnn nnnnnnnnnn nnnnccagaa gatactcatc 1500 ctactcaaat actcnctaac aagacagcaa gattgacacc caccttacag ataaaacaag 1560 ntgtgtgagg cttcactgga ttggtgaact actgatnnag gctttatttc taaatcaaaa 1620 caagacccag tcagactttt atgcctgaaa actttgagga tggtgtacat gcttcaaaag 1680 aacatgtttt aaagacagac ctgacatact cccattttta aaaaaaaaaa aaaaaggtaa 1740 aaaatgagct ggccatggca catgccttta gtctcatctc actgggaggt agaaacaggc 1800 agannnnnnn actcttaagt ttgagaccag cctggttttg tatagggcag ccagggcagc 1860 agtagggtga tctctcaaaa gggggtggaa agataaactt tatctnnnct ccctatcaag 1920 ctatgacttt tatttcatct gaattaaaga cactgaataa tttgagtatt t 1971 74 1951 DNA Homo sapiens misc_feature (1)...(1952) n = A,T,C or G 74 nnnnnnnnnn nnnnnnnnnn nnnnnncgcc cgcgcggcga cgtctccgcg aggcgtcacg 60 gcaccgactg acggccaccc accatggccg cagacncagc gcccgaaggc cgacaccctg 120 gccctgaggc aacggctcat cagctcttcc tgcagactct tttttcccga ggatcctgtt 180 aagattgtcc gggcccaagg gcagtacatg tacgatgaac agggggcaga atacatcgat 240 tgcatcagca atgtggcgca cgttgggcac tgccaccctc tcgtggtcca agcagcacat 300 gagcagaacc aggtgctcaa caccaacagc cggtacctgc atgacaacat cgtggactat 360 gcgcagaggc tgtcagagac cctgccggag cagctctgtg tgttctattt cctgaattct 420 gggtcagaag ccaatgacct ggccctgagg ctggctcgcc actacacggg acaccaggac 480 gtggtggtat tagatcatgc gtatcacggc cacctgagct ccctgattga catcagtccc 540 tacaagttcc gcaacctgga tggccagaag gagtgggtcc acgtggcacc tctcccagac 600 acctaccggg gcccctaccg ggaggaccac cccaacccag ctatggccta tgccaacgag 660 gtgaaacgtg tggtcagcag tgcacaggag aagggcagga agattgcagc cttcttcgct 720 gagtctctgc ccagtgtggg agggcagatc attccccctg ctggctactt ctcccaagtg 780 gcagagcaca tccgcaaggc cggaggggtc tttgttgcag atgagatcca ggttggcttt 840 ggccgggtag gcaagcactt ctgggccttc cagctccagg gaaaagactt cgtccctgac 900 atcgtcacca tgggcaagtc cattggcaac ggccaccctg ttgcctgcgt ggccgcaacc 960 cagcctgtgg cgagggcatt tgaagccacc ggcgttgagt acttcaacac gtttgggggc 1020 agcccagtgt cctgcgctgt ggggctggcc gtcctgaatg tcttggagaa ggagcagctc 1080 caggatcatg ccaccagtgt aggcagcttc ctgatgcagc tcctcgggca gcaaaaaatc 1140 aaacatccca tcgtcgggga tgtcaggggt gttgggctct tcattggtgt ggatctgatc 1200 aaagatgagg ccacaaggac accagcaact gaagaggctg cctacttggt atcaaggctg 1260 aaggagaact acgttttgct gagcactgat ggccctggga ggaacatcct gaagtttaag 1320 cccccaatgt gcttcagcct ggacaatgca cggcaggtgg tggcaaagct ggatgccatt 1380 ctgactgaca tggaagagaa ggtgagaagt tgtgaaacgc tgaggctcca gccctaagcc 1440 agccctgctc tgcctaagtg tactccagaa gaaactcatc tcatccaaat acacgctatt 1500 gagaaggcga gcctgacctc cctcttacag ataaagtcag ctttcagagg ctnnncaggg 1560 tgggggggcn nctgcccgag gccataatgc tannnnnnnn nnnnncccac cccctcctnn 1620 nnncctaacc actgnnngtc tgttgganat nnnnnnaacc cagatgtnnn nnnnnnnnnn 1680 ctgncatccc ctcannnnnn nnnnnnnnnn nnnnagtcag tcaatnnnnn nnnnnnnnnn 1740 nnttcctttc ntgtcnnnnc actgggnggt ggaatggggt agggtgggat actttaaagt 1800 gctnnnnnnn cctgcttaaa taaattagan ccagnnnacc agtnnngtna tttctnnnnn 1860 nnnnnnnnaa agaaaatcct gacatgcaca cccattaaaa atagtacatt tnntacagnt 1920 gaaaaaaaaa aannnaannn nnnnnnnnnn n 1951 75 532 PRT Homo sapiens VARIANT (1)...(9) Xaa = Any amino acid 75 Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Met Phe Ser Lys Leu Ala His 1 5 10 15 Leu Gln Arg Phe Ala Val Leu Ser Arg Gly Val His Ser Ser Val Ala 20 25 30 Ser Ala Thr Ser Val Ala Thr Lys Xaa Xaa Xaa Xaa Xaa Xaa Lys Thr 35 40 45 Val Gln Gly Pro Pro Thr Ser Asp Asp Ile Phe Glu Arg Glu Tyr Lys 50 55 60 Tyr Gly Ala His Asn Tyr His Pro Leu Pro Val Ala Leu Glu Arg Gly 65 70 75 80 Lys Gly Ile Tyr Leu Trp Asp Val Glu Gly Arg Lys Tyr Phe Asp Phe 85 90 95 Leu Ser Ser Tyr Ser Ala Val Asn Gln Gly His Cys His Pro Lys Ile 100 105 110 Val Asn Ala Leu Lys Ser Gln Val Asp Lys Leu Thr Leu Thr Ser Arg 115 120 125 Xaa Xaa Xaa Xaa Xaa Xaa Ala Phe Tyr Asn Asn Val Leu Gly Glu Tyr 130 135 140 Glu Glu Tyr Ile Thr Lys Leu Phe Asn Tyr His Lys Val Leu Pro Xaa 145 150 155 160 Met Asn Thr Gly Val Glu Ala Gly Glu Thr Ala Cys Lys Leu Ala Arg 165 170 175 Lys Trp Gly Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Tyr Thr Val Lys 180 185 190 Gly Ile Gln Lys Tyr Lys Ala Lys Ile Val Phe Ala Ala Gly Asn Xaa 195 200 205 Xaa Xaa Xaa Xaa Phe Trp Gly Arg Thr Leu Ser Ala Ile Ser Ser Ser 210 215 220 Thr Asp Pro Thr Ser Tyr Asp Gly Xaa Phe Gly Pro Phe Met Pro Gly 225 230 235 240 Phe Asp Ile Ile Pro Tyr Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 245 250 255 Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Asn Asp 260 265 270 Leu Pro Ala Leu Glu Arg Ala Leu Gln Asp Pro Xaa Xaa Asn Val Ala 275 280 285 Ala Phe Met Val Glu Pro Ile Gln Gly Glu Ala Gly Val Val Val Pro 290 295 300 Asp Pro Gly Tyr Leu Met Gly Val Arg Glu Leu Cys Thr Arg His Gln 305 310 315 320 Val Leu Phe Ile Ala Asp Glu Ile Gln Thr Gly Leu Ala Arg Thr Gly 325 330 335 Arg Xaa Xaa Xaa Trp Leu Ala Val Asp Tyr Glu Asn Val Arg Pro Asp 340 345 350 Ile Val Leu Leu Gly Lys Ala Leu Ser Gly Gly Leu Tyr Pro Val Ser 355 360 365 Ala Val Leu Cys Asp Asp Asp Ile Met Leu Thr Ile Lys Pro Gly Xaa 370 375 380 Xaa Glu His Gly Ser Thr Tyr Gly Gly Asn Pro Leu Gly Cys Arg Val 385 390 395 400 Ala Ile Ala Ala Leu Glu Val Leu Glu Glu Glu Asn Leu Ala Glu Asn 405 410 415 Ala Asp Lys Leu Gly Xaa Ile Ile Leu Arg Asn Glu Leu Xaa Xaa Met 420 425 430 Lys Leu Pro Ser Asp Val Val Thr Ala Val Arg Gly Lys Gly Leu Leu 435 440 445 Asn Xaa Xaa Xaa Xaa Xaa Xaa Ala Ile Val Ile Lys Glu Thr Lys Asp 450 455 460 Trp Asp Ala Trp Lys Val Cys Leu Arg Leu Arg Asp Asn Gly Leu Leu 465 470 475 480 Ala Lys Pro Thr His Gly Xaa Xaa Asp Ile Ile Arg Phe Ala Pro Pro 485 490 495 Leu Val Ile Lys Glu Asp Glu Leu Arg Glu Ser Ile Glu Ile Ile Asn 500 505 510 Lys Thr Ile Leu Ser Phe Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 515 520 525 Xaa Xaa Xaa Xaa 530 76 532 PRT Homo sapiens VARIANT (378)...(386) Xaa = Any amino acid 76 Met Ala Ser Met Leu Leu Ala Gln Arg Leu Ala Cys Ser Phe Gln His 1 5 10 15 Thr Tyr Arg Leu Leu Val Pro Gly Ser Arg His Ile Ser Gln Ala Ala 20 25 30 Ala Lys Val Asp Val Glu Phe Asp Tyr Asp Gly Pro Leu Met Lys Thr 35 40 45 Glu Val Pro Gly Pro Arg Ser Gln Glu Leu Met Lys Gln Leu Asn Ile 50 55 60 Ile Gln Asn Ala Glu Ala Val His Phe Phe Cys Asn Tyr Glu Glu Ser 65 70 75 80 Arg Gly Asn Tyr Leu Val Asp Val Asp Gly Asn Arg Met Leu Asp Leu 85 90 95 Tyr Ser Gln Ile Ser Ser Val Pro Ile Gly Tyr Ser Asp Pro Ala Leu 100 105 110 Val Lys Leu Ile Gln Gln Pro Gln Asn Ala Ser Met Phe Val Asn Arg 115 120 125 Pro Ala Leu Glu Ile Leu Pro Pro Glu Asn Phe Val Glu Lys Leu Arg 130 135 140 Gln Ser Leu Leu Ser Val Ala Pro Lys Gly Met Ser Gln Leu Ile Thr 145 150 155 160 Met Ala Cys Gly Ser Cys Ser Asn Glu Asn Ala Leu Lys Thr Ile Phe 165 170 175 Met Trp Tyr Arg Ser Lys Glu Arg Gly Gln Arg Gly Phe Ser Lys Glu 180 185 190 Glu Leu Glu Thr Cys Met Ile Asn Gln Ala Pro Trp Cys Pro Asp Tyr 195 200 205 Ser Ile Leu Ser Phe Met Gly Ser Phe His Gly Arg Thr Met Gly Cys 210 215 220 Leu Ala Thr Thr His Ser Lys Ala Ile His Lys Ile Asp Ile Pro Ser 225 230 235 240 Phe Asp Trp Pro Ile Ala Pro Phe Pro Arg Leu Lys Tyr Pro Leu Glu 245 250 255 Glu Phe Val Lys Glu Asn Gln Gln Glu Glu Ala Gly Cys Leu Glu Glu 260 265 270 Val Glu Asp Leu Ile Val Lys Tyr Arg Lys Lys Lys Lys Thr Val Ala 275 280 285 Gly Ile Ile Val Glu Pro Ile Gln Ser Glu Gly Gly Asp Asn His Ala 290 295 300 Ser Asp Asp Phe Phe Arg Lys Leu Arg Asp Ile Ala Arg Lys His Cys 305 310 315 320 Cys Ala Phe Leu Val Asp Glu Val Gln Thr Gly Gly Gly Cys Thr Gly 325 330 335 Lys Phe Trp Ala His Glu His Trp Gly Leu Asp Asp Pro Ala Asp Val 340 345 350 Met Thr Phe Ser Lys Lys Met Met Thr Gly Gly Phe Phe Leu Lys Glu 355 360 365 Glu Phe Arg Pro Asn Ala Pro Tyr Arg Xaa Xaa Xaa Xaa Xaa Xaa Xaa 370 375 380 Xaa Xaa Ile Phe Asn Thr Trp Leu Gly Asp Pro Ser Lys Asn Leu Leu 385 390 395 400 Leu Ala Glu Val Ile Asn Ile Ile Lys Arg Glu Asp Leu Leu Asn Asn 405 410 415 Ala Ala His Ala Gly Lys Ala Leu Leu Thr Gly Leu Leu Asp Leu Gln 420 425 430 Ala Arg Tyr Pro Gln Phe Ile Ser Arg Val Arg Gly Arg Gly Thr Phe 435 440 445 Cys Ser Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Phe Asp Thr Pro Asp Asp 450 455 460 Ser Ile Arg Asn Lys Leu Ile Leu Ile Ala Arg Asn Lys Gly Val Val 465 470 475 480 Leu Gly Gly Cys Gly Asp Xaa Xaa Lys Ser Ile Arg Phe Arg Pro Thr 485 490 495 Leu Val Phe Arg Asp His His Ala His Leu Phe Leu Asn Ile Phe Ser 500 505 510 Asp Ile Leu Ala Asp Phe Lys Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 515 520 525 Xaa Xaa Xaa Xaa 530 77 27 PRT Artificial Sequence Aminotransferase class III pyridoxal-phosphate attachment site consensus sequence 77 Xaa Xaa Xaa Asp Glu Xaa Xaa Xaa Gly Xaa Xaa Xaa Xaa Xaa Xaa Asp 1 5 10 15 Xaa Xaa Xaa Xaa Xaa Lys Xaa Xaa Xaa Xaa Xaa 20 25 78 1687 DNA Homo sapiens CDS (160)...(1470) misc_feature (1)...(1687) n = A,T,C or G 78 cccacgcgtn cgggcatgtg ggagccacat gctgggtgcc ccagacagcc taatgctcat 60 tctcaggccg ggctttccag cctctaggtg ctgtgctgtc ctgaggcctg ggccatggtg 120 cccaaggaaa gcccctgaag ctcaccagga ggaagaagc atg cag ggc act cct 174 Met Gln Gly Thr Pro 1 5 gga ggc ggg acg cgc cct ggg cca tcc ccc gtg gac agg cgg acg ctc 222 Gly Gly Gly Thr Arg Pro Gly Pro Ser Pro Val Asp Arg Arg Thr Leu 10 15 20 ctg gtc ttc agc ttt atc ctg gca gca gct ttg ggc caa atg aat ttc 270 Leu Val Phe Ser Phe Ile Leu Ala Ala Ala Leu Gly Gln Met Asn Phe 25 30 35 aca ggg gac cag gtt ctt cga gtc ctg gcc aaa gat gag aag cag ctt 318 Thr Gly Asp Gln Val Leu Arg Val Leu Ala Lys Asp Glu Lys Gln Leu 40 45 50 tca ctt ctc ggg gat ctg gag ggc ctg aaa ccc cag aag gtg gac ttc 366 Ser Leu Leu Gly Asp Leu Glu Gly Leu Lys Pro Gln Lys Val Asp Phe 55 60 65 tgg cgt ggc cca gcc agg ccc agc ctc cct gtg gat atg aga gtt cct 414 Trp Arg Gly Pro Ala Arg Pro Ser Leu Pro Val Asp Met Arg Val Pro 70 75 80 85 ttc tcc gaa ctg aaa gac atc aaa gct tat ctg gag tct cat gga ctt 462 Phe Ser Glu Leu Lys Asp Ile Lys Ala Tyr Leu Glu Ser His Gly Leu 90 95 100 gct tac agc atc atg ata aag gac atc cag gtg ctg ctg gat gag gaa 510 Ala Tyr Ser Ile Met Ile Lys Asp Ile Gln Val Leu Leu Asp Glu Glu 105 110 115 aga cag gcc atg gcg aaa tcc cgc cgg ctg gag cgc agc acc aac agc 558 Arg Gln Ala Met Ala Lys Ser Arg Arg Leu Glu Arg Ser Thr Asn Ser 120 125 130 ttc agt tac tca tca tac cac acc ctg gag gag ata tat agc tgg att 606 Phe Ser Tyr Ser Ser Tyr His Thr Leu Glu Glu Ile Tyr Ser Trp Ile 135 140 145 gac aac ttt gta atg gag cat tcc gat att gtc tca aaa att cag att 654 Asp Asn Phe Val Met Glu His Ser Asp Ile Val Ser Lys Ile Gln Ile 150 155 160 165 ggc aac agc ttt gaa aac cag tcc att ctt gtc ctg aag ttc agc act 702 Gly Asn Ser Phe Glu Asn Gln Ser Ile Leu Val Leu Lys Phe Ser Thr 170 175 180 gga ggt tct cgg cac cca gcc atc tgg atc gac act gga att cac tcc 750 Gly Gly Ser Arg His Pro Ala Ile Trp Ile Asp Thr Gly Ile His Ser 185 190 195 cgg gag tgg atc acc cat gcc acc ggc atc tgg act gcc aat aag att 798 Arg Glu Trp Ile Thr His Ala Thr Gly Ile Trp Thr Ala Asn Lys Ile 200 205 210 gtc agt gat tat ggc aaa gac cgt gtc ctg aca gac ata ctg aat gcc 846 Val Ser Asp Tyr Gly Lys Asp Arg Val Leu Thr Asp Ile Leu Asn Ala 215 220 225 atg gac atc ttc ata gag ctc gtc aca aac cct gat ggg ttt gct ttt 894 Met Asp Ile Phe Ile Glu Leu Val Thr Asn Pro Asp Gly Phe Ala Phe 230 235 240 245 acc cac agc atg aac cgc tta tgg cgg aag aac aag tcc atc aga cct 942 Thr His Ser Met Asn Arg Leu Trp Arg Lys Asn Lys Ser Ile Arg Pro 250 255 260 gga atc ttc tgc atc ggc gtg gat ctc aac agg aac tgg aag tcg ggt 990 Gly Ile Phe Cys Ile Gly Val Asp Leu Asn Arg Asn Trp Lys Ser Gly 265 270 275 ttt gga gga aat ggt tct aac agc aac ccc tgc tca gaa act tat cac 1038 Phe Gly Gly Asn Gly Ser Asn Ser Asn Pro Cys Ser Glu Thr Tyr His 280 285 290 ggg ccc tcc cct cag tcg gag tcg gag gtg gct gcc ata gtg aac ttc 1086 Gly Pro Ser Pro Gln Ser Glu Ser Glu Val Ala Ala Ile Val Asn Phe 295 300 305 atc aca gcc cat ggc aac ttc aag gct ctg atc tcc atc cac agc tac 1134 Ile Thr Ala His Gly Asn Phe Lys Ala Leu Ile Ser Ile His Ser Tyr 310 315 320 325 tct cag atg ctt atg tac cct tac ggc cga ttg ctg gag ccc gtt tca 1182 Ser Gln Met Leu Met Tyr Pro Tyr Gly Arg Leu Leu Glu Pro Val Ser 330 335 340 aat cag agg gag ttg tac gat ctt gcc aag gat gcg gtg gag gcc ttg 1230 Asn Gln Arg Glu Leu Tyr Asp Leu Ala Lys Asp Ala Val Glu Ala Leu 345 350 355 tat aag gtc cat ggg atc gag tac att ttt ggc agc atc agc acc acc 1278 Tyr Lys Val His Gly Ile Glu Tyr Ile Phe Gly Ser Ile Ser Thr Thr 360 365 370 ctc tat gtg gcc agt ggg atc acc gtc gac tgg gcc tat gac agt ggc 1326 Leu Tyr Val Ala Ser Gly Ile Thr Val Asp Trp Ala Tyr Asp Ser Gly 375 380 385 atc aag tac gcc ttc agc ttt gag ctc cgg gac act ggg cag tat ggc 1374 Ile Lys Tyr Ala Phe Ser Phe Glu Leu Arg Asp Thr Gly Gln Tyr Gly 390 395 400 405 ttc ctg ctg ccg gcc aca cag atc atc ccc acg gcc cag gag acg tgg 1422 Phe Leu Leu Pro Ala Thr Gln Ile Ile Pro Thr Ala Gln Glu Thr Trp 410 415 420 atg gcg ctt cgg acc atc atg gag cac acc ctg aat cac ccc tac tag 1470 Met Ala Leu Arg Thr Ile Met Glu His Thr Leu Asn His Pro Tyr * 425 430 435 cagcacgact gagggcagga ggctccatcc ttctccccaa ggtctgtggc tcctcccgaa 1530 acccaagtta tgcatcccca tccccatgcc ctcatcccga cctcttagaa aataaataca 1590 agtttgaaca ggcaaaaaaa aaaaaaaaaa aaaaaattgg cggccgcaag cttattcctt 1650 taagtgaggg ttaattttag cttggcactg gccgncg 1687 79 436 PRT Homo sapiens 79 Met Gln Gly Thr Pro Gly Gly Gly Thr Arg Pro Gly Pro Ser Pro Val 1 5 10 15 Asp Arg Arg Thr Leu Leu Val Phe Ser Phe Ile Leu Ala Ala Ala Leu 20 25 30 Gly Gln Met Asn Phe Thr Gly Asp Gln Val Leu Arg Val Leu Ala Lys 35 40 45 Asp Glu Lys Gln Leu Ser Leu Leu Gly Asp Leu Glu Gly Leu Lys Pro 50 55 60 Gln Lys Val Asp Phe Trp Arg Gly Pro Ala Arg Pro Ser Leu Pro Val 65 70 75 80 Asp Met Arg Val Pro Phe Ser Glu Leu Lys Asp Ile Lys Ala Tyr Leu 85 90 95 Glu Ser His Gly Leu Ala Tyr Ser Ile Met Ile Lys Asp Ile Gln Val 100 105 110 Leu Leu Asp Glu Glu Arg Gln Ala Met Ala Lys Ser Arg Arg Leu Glu 115 120 125 Arg Ser Thr Asn Ser Phe Ser Tyr Ser Ser Tyr His Thr Leu Glu Glu 130 135 140 Ile Tyr Ser Trp Ile Asp Asn Phe Val Met Glu His Ser Asp Ile Val 145 150 155 160 Ser Lys Ile Gln Ile Gly Asn Ser Phe Glu Asn Gln Ser Ile Leu Val 165 170 175 Leu Lys Phe Ser Thr Gly Gly Ser Arg His Pro Ala Ile Trp Ile Asp 180 185 190 Thr Gly Ile His Ser Arg Glu Trp Ile Thr His Ala Thr Gly Ile Trp 195 200 205 Thr Ala Asn Lys Ile Val Ser Asp Tyr Gly Lys Asp Arg Val Leu Thr 210 215 220 Asp Ile Leu Asn Ala Met Asp Ile Phe Ile Glu Leu Val Thr Asn Pro 225 230 235 240 Asp Gly Phe Ala Phe Thr His Ser Met Asn Arg Leu Trp Arg Lys Asn 245 250 255 Lys Ser Ile Arg Pro Gly Ile Phe Cys Ile Gly Val Asp Leu Asn Arg 260 265 270 Asn Trp Lys Ser Gly Phe Gly Gly Asn Gly Ser Asn Ser Asn Pro Cys 275 280 285 Ser Glu Thr Tyr His Gly Pro Ser Pro Gln Ser Glu Ser Glu Val Ala 290 295 300 Ala Ile Val Asn Phe Ile Thr Ala His Gly Asn Phe Lys Ala Leu Ile 305 310 315 320 Ser Ile His Ser Tyr Ser Gln Met Leu Met Tyr Pro Tyr Gly Arg Leu 325 330 335 Leu Glu Pro Val Ser Asn Gln Arg Glu Leu Tyr Asp Leu Ala Lys Asp 340 345 350 Ala Val Glu Ala Leu Tyr Lys Val His Gly Ile Glu Tyr Ile Phe Gly 355 360 365 Ser Ile Ser Thr Thr Leu Tyr Val Ala Ser Gly Ile Thr Val Asp Trp 370 375 380 Ala Tyr Asp Ser Gly Ile Lys Tyr Ala Phe Ser Phe Glu Leu Arg Asp 385 390 395 400 Thr Gly Gln Tyr Gly Phe Leu Leu Pro Ala Thr Gln Ile Ile Pro Thr 405 410 415 Ala Gln Glu Thr Trp Met Ala Leu Arg Thr Ile Met Glu His Thr Leu 420 425 430 Asn His Pro Tyr 435 80 1311 DNA Homo sapiens CDS (1)...(1311) 80 atg cag ggc act cct gga ggc ggg acg cgc cct ggg cca tcc ccc gtg 48 Met Gln Gly Thr Pro Gly Gly Gly Thr Arg Pro Gly Pro Ser Pro Val 1 5 10 15 gac agg cgg acg ctc ctg gtc ttc agc ttt atc ctg gca gca gct ttg 96 Asp Arg Arg Thr Leu Leu Val Phe Ser Phe Ile Leu Ala Ala Ala Leu 20 25 30 ggc caa atg aat ttc aca ggg gac cag gtt ctt cga gtc ctg gcc aaa 144 Gly Gln Met Asn Phe Thr Gly Asp Gln Val Leu Arg Val Leu Ala Lys 35 40 45 gat gag aag cag ctt tca ctt ctc ggg gat ctg gag ggc ctg aaa ccc 192 Asp Glu Lys Gln Leu Ser Leu Leu Gly Asp Leu Glu Gly Leu Lys Pro 50 55 60 cag aag gtg gac ttc tgg cgt ggc cca gcc agg ccc agc ctc cct gtg 240 Gln Lys Val Asp Phe Trp Arg Gly Pro Ala Arg Pro Ser Leu Pro Val 65 70 75 80 gat atg aga gtt cct ttc tcc gaa ctg aaa gac atc aaa gct tat ctg 288 Asp Met Arg Val Pro Phe Ser Glu Leu Lys Asp Ile Lys Ala Tyr Leu 85 90 95 gag tct cat gga ctt gct tac agc atc atg ata aag gac atc cag gtg 336 Glu Ser His Gly Leu Ala Tyr Ser Ile Met Ile Lys Asp Ile Gln Val 100 105 110 ctg ctg gat gag gaa aga cag gcc atg gcg aaa tcc cgc cgg ctg gag 384 Leu Leu Asp Glu Glu Arg Gln Ala Met Ala Lys Ser Arg Arg Leu Glu 115 120 125 cgc agc acc aac agc ttc agt tac tca tca tac cac acc ctg gag gag 432 Arg Ser Thr Asn Ser Phe Ser Tyr Ser Ser Tyr His Thr Leu Glu Glu 130 135 140 ata tat agc tgg att gac aac ttt gta atg gag cat tcc gat att gtc 480 Ile Tyr Ser Trp Ile Asp Asn Phe Val Met Glu His Ser Asp Ile Val 145 150 155 160 tca aaa att cag att ggc aac agc ttt gaa aac cag tcc att ctt gtc 528 Ser Lys Ile Gln Ile Gly Asn Ser Phe Glu Asn Gln Ser Ile Leu Val 165 170 175 ctg aag ttc agc act gga ggt tct cgg cac cca gcc atc tgg atc gac 576 Leu Lys Phe Ser Thr Gly Gly Ser Arg His Pro Ala Ile Trp Ile Asp 180 185 190 act gga att cac tcc cgg gag tgg atc acc cat gcc acc ggc atc tgg 624 Thr Gly Ile His Ser Arg Glu Trp Ile Thr His Ala Thr Gly Ile Trp 195 200 205 act gcc aat aag att gtc agt gat tat ggc aaa gac cgt gtc ctg aca 672 Thr Ala Asn Lys Ile Val Ser Asp Tyr Gly Lys Asp Arg Val Leu Thr 210 215 220 gac ata ctg aat gcc atg gac atc ttc ata gag ctc gtc aca aac cct 720 Asp Ile Leu Asn Ala Met Asp Ile Phe Ile Glu Leu Val Thr Asn Pro 225 230 235 240 gat ggg ttt gct ttt acc cac agc atg aac cgc tta tgg cgg aag aac 768 Asp Gly Phe Ala Phe Thr His Ser Met Asn Arg Leu Trp Arg Lys Asn 245 250 255 aag tcc atc aga cct gga atc ttc tgc atc ggc gtg gat ctc aac agg 816 Lys Ser Ile Arg Pro Gly Ile Phe Cys Ile Gly Val Asp Leu Asn Arg 260 265 270 aac tgg aag tcg ggt ttt gga gga aat ggt tct aac agc aac ccc tgc 864 Asn Trp Lys Ser Gly Phe Gly Gly Asn Gly Ser Asn Ser Asn Pro Cys 275 280 285 tca gaa act tat cac ggg ccc tcc cct cag tcg gag tcg gag gtg gct 912 Ser Glu Thr Tyr His Gly Pro Ser Pro Gln Ser Glu Ser Glu Val Ala 290 295 300 gcc ata gtg aac ttc atc aca gcc cat ggc aac ttc aag gct ctg atc 960 Ala Ile Val Asn Phe Ile Thr Ala His Gly Asn Phe Lys Ala Leu Ile 305 310 315 320 tcc atc cac agc tac tct cag atg ctt atg tac cct tac ggc cga ttg 1008 Ser Ile His Ser Tyr Ser Gln Met Leu Met Tyr Pro Tyr Gly Arg Leu 325 330 335 ctg gag ccc gtt tca aat cag agg gag ttg tac gat ctt gcc aag gat 1056 Leu Glu Pro Val Ser Asn Gln Arg Glu Leu Tyr Asp Leu Ala Lys Asp 340 345 350 gcg gtg gag gcc ttg tat aag gtc cat ggg atc gag tac att ttt ggc 1104 Ala Val Glu Ala Leu Tyr Lys Val His Gly Ile Glu Tyr Ile Phe Gly 355 360 365 agc atc agc acc acc ctc tat gtg gcc agt ggg atc acc gtc gac tgg 1152 Ser Ile Ser Thr Thr Leu Tyr Val Ala Ser Gly Ile Thr Val Asp Trp 370 375 380 gcc tat gac agt ggc atc aag tac gcc ttc agc ttt gag ctc cgg gac 1200 Ala Tyr Asp Ser Gly Ile Lys Tyr Ala Phe Ser Phe Glu Leu Arg Asp 385 390 395 400 act ggg cag tat ggc ttc ctg ctg ccg gcc aca cag atc atc ccc acg 1248 Thr Gly Gln Tyr Gly Phe Leu Leu Pro Ala Thr Gln Ile Ile Pro Thr 405 410 415 gcc cag gag acg tgg atg gcg ctt cgg acc atc atg gag cac acc ctg 1296 Ala Gln Glu Thr Trp Met Ala Leu Arg Thr Ile Met Glu His Thr Leu 420 425 430 aat cac ccc tac tag 1311 Asn His Pro Tyr * 435 81 304 PRT Artificial Sequence Amino acid consensus sequence 81 Tyr His Asn Leu Glu Glu Ile Tyr Ala Trp Leu Asp Leu Leu Val Ser 1 5 10 15 Asn Phe Pro Asp Leu Val Ser Lys Val Ser Ile Gly Lys Ser Tyr Glu 20 25 30 Gly Arg Asp Leu Lys Val Leu Lys Ile Ser Asp Asn Pro Ala Thr Gly 35 40 45 Glu Asn Glu Pro Glu Val Phe Ala Val Ala Gly Trp Ile His Ala Arg 50 55 60 Glu Trp Val Thr Ser Ala Thr Leu Leu Trp Leu Leu Lys Glu Leu Val 65 70 75 80 Ala Asn Tyr Gly Ser Asp Lys Thr Ile Thr Lys Leu Leu Asp Gly Leu 85 90 95 Asp Leu Phe Tyr Ile Leu Pro Val Phe Asn Pro Asp Gly Tyr Ala Tyr 100 105 110 Ser Ile Thr Thr Asp Ser Tyr Arg Met Trp Arg Lys Thr Arg Ser Pro 115 120 125 Asn Ala Gly Ser Phe Cys Val Gly Thr Asp Pro Asn Arg Asn Trp Tyr 130 135 140 Ala Gln Trp Gly Gly Met Gly Ala Ser Ser Tyr Ser Pro Cys Ser Glu 145 150 155 160 Thr Tyr Glu Gly Thr Ala Pro Phe Ser Glu Pro Glu Thr Lys Ala Val 165 170 175 Glu Asp Phe Ile Arg Ser Trp Leu Gly Gly Gly Lys Gln Asn Ile Lys 180 185 190 Ala Tyr Ile Thr Phe His Ser Tyr Ser Gln Leu Leu Leu Tyr Pro Tyr 195 200 205 Gly Tyr Asp Tyr Asn Leu Asn Pro Asp Ala Asn Asp Leu Asp Glu Leu 210 215 220 Ser Asp Leu Lys Ile Ala Ala Asp Ala Leu Ser Ala Arg His Gly Thr 225 230 235 240 Tyr Tyr Thr Leu Gly Leu Pro Gly Ser Ser Thr Ile Tyr Pro Ala Ser 245 250 255 Ala Gly Gly Ser Asp Asp Trp Ala Tyr Asp Val Gly Ile Ile Lys Tyr 260 265 270 Ala Phe Thr Phe Glu Leu Arg Pro Asp Thr Gly Ser Tyr Gly Asn Pro 275 280 285 Cys Phe Leu Leu Pro Glu Glu Gln Ile Ile Pro Thr Gly Ser Glu Glu 290 295 300 82 324 PRT Artificial Sequence Amino acid consensus sequence 82 Tyr His Ser Tyr Glu Glu Ile Asn Ala Trp Leu Asp Asp Leu Ala Arg 1 5 10 15 Asn Tyr Pro Asp Leu Thr Ser Val Ser Leu Ile Ser Ile Gly Lys Ser 20 25 30 Tyr Glu Gly Arg Pro Ile Lys Val Leu Lys Ile Lys Pro Ala Val Phe 35 40 45 Ile Asp Ala Gly Ile His Ala Arg Glu Trp Ile Ala Pro Ala Thr Ala 50 55 60 Leu Tyr Leu Ile Asn Gln Leu Leu Thr Asn Glu Thr Glu Tyr Ser Lys 65 70 75 80 Asp Pro Asp Asp Glu Gly Ser Val Thr Lys Leu Leu Asp Lys Leu Asp 85 90 95 Trp Tyr Ile Val Pro Val Met Asn Pro Asp Gly Tyr Glu Tyr Thr His 100 105 110 Thr Ser Thr Asp Arg Leu Trp Arg Lys Asn Arg Ser Pro Asn Gly Ala 115 120 125 Ser Gly Ser Gln Gly Thr Trp Tyr Asn Cys Tyr Gly Val Asp Leu Asn 130 135 140 Arg Asn Phe Asp Phe His Asn Trp Gly Glu Ile Gly Gly Ser Ser Ser 145 150 155 160 Leu Pro Cys Ser Glu Thr Tyr Ala Gly Ser Ser Pro Phe Ser Glu Trp 165 170 175 Glu Pro Glu Thr Lys Ala Leu Leu Asp Phe Ile Leu Ser Asn Glu Ile 180 185 190 Gly Lys Gly Arg Ile Lys Ala Tyr Ile Ser Leu His Ser Tyr Ser Gln 195 200 205 Leu Leu Leu Tyr Pro Tyr Gly Tyr Thr Asn Ala Thr Val Pro Pro Asn 210 215 220 Gly Glu Asp Leu His Lys Glu Val Ala Lys Ala Ala Ala Lys Ala Ile 225 230 235 240 Gly Asp Tyr Tyr Phe Gly Gly Thr Leu Tyr Thr Pro Gly Ser Ser Ser 245 250 255 Ala Asp Pro Asp Leu Asp Ile Thr Leu Tyr Pro Ala Ser Gly Gly Ser 260 265 270 Asp Asp Trp Ala Tyr Gly Thr Leu Lys Gly Val Lys Tyr Ser Tyr Thr 275 280 285 Ile Glu Leu Arg Asp Thr Gly Asp Asp Ala Gly Arg Tyr Gly Phe Leu 290 295 300 Leu Pro Pro Ser Cys Val Lys Pro Val Arg Met Glu Gln Ile Ile Pro 305 310 315 320 Thr Gly Glu Glu 83 82 PRT Artificial Sequence Amino acid consensus sequence 83 Gln Val Leu Arg Val Lys Val Ala Asp Glu Asp Gln Val Lys Leu Leu 1 5 10 15 Lys Asp Leu Glu Asn Thr Glu His Leu Glu Leu Asp Phe Trp Lys Pro 20 25 30 Asp Ser Ala Thr Pro Ile Lys Pro Gly Ser Thr Val Asp Phe Arg Val 35 40 45 Pro Ala Glu Asp Ile Gln Ala Val Lys Ser Phe Leu Glu Gln Ser Gly 50 55 60 Ile His Tyr Glu Val Leu Ile Glu Asp Val Gln Glu Leu Leu Glu Glu 65 70 75 80 Gln Phe 84 23 PRT Artificial Sequence Zinc carboxypeptidase zinc-binding region 1 signature consensus sequence 84 Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa His Xaa Xaa Glu Xaa Xaa Xaa 1 5 10 15 Xaa Xaa Xaa Xaa Xaa Xaa Xaa 20 85 11 PRT Artificial Sequence Zinc carboxypeptidase zinc-binding region 2 signature consensus sequence 85 His Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Pro Xaa 1 5 10 86 22 PRT Artificial Sequence Zinc carboxypeptidase zinc-binding region 1 signature 86 Pro Ala Ile Trp Ile Asp Thr Gly His Ser Arg Glu Trp Ile Thr His 1 5 10 15 Ala Thr Gly Ile Trp Thr 20 87 11 PRT Artificial Sequence Zinc carboxypeptidase zinc-binding region 2 signature 87 His Ser Tyr Ser Gln Met Leu Met Tyr Pro Tyr 1 5 10 88 5389 DNA Homo sapiens CDS (6)...(3638) 88 ttggt atg gca tca cag ctg caa gtg ttt tcg ccc cca tca gtg tcg tcg 50 Met Ala Ser Gln Leu Gln Val Phe Ser Pro Pro Ser Val Ser Ser 1 5 10 15 agt gcc ttc tgc agt gcg aag aaa ctg aaa ata gag ccc tct ggc tgg 98 Ser Ala Phe Cys Ser Ala Lys Lys Leu Lys Ile Glu Pro Ser Gly Trp 20 25 30 gat gtt tca gga cag agt agc aac gac aaa tat tat acc cac agc aaa 146 Asp Val Ser Gly Gln Ser Ser Asn Asp Lys Tyr Tyr Thr His Ser Lys 35 40 45 acc ctc cca gcc aca caa ggg caa gcc aac tcc tct cac cag gta gca 194 Thr Leu Pro Ala Thr Gln Gly Gln Ala Asn Ser Ser His Gln Val Ala 50 55 60 aat ttc aac atc cct gct tac gac cag ggc ctc ctc ctc cca gct cct 242 Asn Phe Asn Ile Pro Ala Tyr Asp Gln Gly Leu Leu Leu Pro Ala Pro 65 70 75 gca gtg gag cat att gtt gta aca gcc gct gat agc tcg ggc agt gct 290 Ala Val Glu His Ile Val Val Thr Ala Ala Asp Ser Ser Gly Ser Ala 80 85 90 95 gct aca tca acc ttc caa agc agc cag acc ctg act ccc aga agc aac 338 Ala Thr Ser Thr Phe Gln Ser Ser Gln Thr Leu Thr Pro Arg Ser Asn 100 105 110 gtt tct ttg ctt gag cca tat caa aaa tgt gga ttg aaa cga aaa agt 386 Val Ser Leu Leu Glu Pro Tyr Gln Lys Cys Gly Leu Lys Arg Lys Ser 115 120 125 gag gaa gtt gac agc aac ggt agt gtg cag atc ata gaa gaa cat ccc 434 Glu Glu Val Asp Ser Asn Gly Ser Val Gln Ile Ile Glu Glu His Pro 130 135 140 cct ctc atg ctg caa aac agg act gtg gtg ggt gct gct gcc aca acc 482 Pro Leu Met Leu Gln Asn Arg Thr Val Val Gly Ala Ala Ala Thr Thr 145 150 155 acc act gtg acc aca aag agt agc agt tcc agc gga gaa ggg gat tac 530 Thr Thr Val Thr Thr Lys Ser Ser Ser Ser Ser Gly Glu Gly Asp Tyr 160 165 170 175 cag ctg gtc cag cat gag atc ctt tgc tct atg acc aat agc tat gaa 578 Gln Leu Val Gln His Glu Ile Leu Cys Ser Met Thr Asn Ser Tyr Glu 180 185 190 gtc ttg gag ttc cta ggc cgg ggg aca ttt gga cag gtg gct aag tgc 626 Val Leu Glu Phe Leu Gly Arg Gly Thr Phe Gly Gln Val Ala Lys Cys 195 200 205 tgg aag agg agc acc aag gaa att gtg gct att aaa atc ttg aag aac 674 Trp Lys Arg Ser Thr Lys Glu Ile Val Ala Ile Lys Ile Leu Lys Asn 210 215 220 cac ccc tcc tat gcc aga caa gga cag att gaa gtg agc atc ctt tcc 722 His Pro Ser Tyr Ala Arg Gln Gly Gln Ile Glu Val Ser Ile Leu Ser 225 230 235 cgc cta agc agt gaa aat gct gat gag tat aat ttt gtc cgt tca tac 770 Arg Leu Ser Ser Glu Asn Ala Asp Glu Tyr Asn Phe Val Arg Ser Tyr 240 245 250 255 gag tgc ttt cag cat aag aat cac acc tgc ctt gtt ttt gaa atg ttg 818 Glu Cys Phe Gln His Lys Asn His Thr Cys Leu Val Phe Glu Met Leu 260 265 270 gag cag aac tta tat gat ttt cta aag caa aac aaa ttt agc cca ctg 866 Glu Gln Asn Leu Tyr Asp Phe Leu Lys Gln Asn Lys Phe Ser Pro Leu 275 280 285 cca ctc aag tac atc aga cca atc ttg cag cag gtg gcc aca gcc ttg 914 Pro Leu Lys Tyr Ile Arg Pro Ile Leu Gln Gln Val Ala Thr Ala Leu 290 295 300 atg aag ctc aag agt ctt ggt ctg atc cac gct gac ctt aag cct gaa 962 Met Lys Leu Lys Ser Leu Gly Leu Ile His Ala Asp Leu Lys Pro Glu 305 310 315 aac atc atg ctg gtt gat cca gtt cgc cag ccc tac cga gtg aag gtc 1010 Asn Ile Met Leu Val Asp Pro Val Arg Gln Pro Tyr Arg Val Lys Val 320 325 330 335 ttt gac ttt ggt tct gct agt cac gtt tcc aaa gct gtg tgc tca acc 1058 Phe Asp Phe Gly Ser Ala Ser His Val Ser Lys Ala Val Cys Ser Thr 340 345 350 tac tta cag tca cgt tac tac aga gct cct gaa att att ctt ggg tta 1106 Tyr Leu Gln Ser Arg Tyr Tyr Arg Ala Pro Glu Ile Ile Leu Gly Leu 355 360 365 cca ttt tgt gaa gct att gat atg tgg tca ctg ggc tgt gtg ata gct 1154 Pro Phe Cys Glu Ala Ile Asp Met Trp Ser Leu Gly Cys Val Ile Ala 370 375 380 gag ctg ttc ctg gga tgg cct ctt tat cct ggt gct tca gaa tat gat 1202 Glu Leu Phe Leu Gly Trp Pro Leu Tyr Pro Gly Ala Ser Glu Tyr Asp 385 390 395 cag att cgt tat att tca caa aca caa ggc ttg cca gct gaa tat ctt 1250 Gln Ile Arg Tyr Ile Ser Gln Thr Gln Gly Leu Pro Ala Glu Tyr Leu 400 405 410 415 ctc agt gcc gga aca aaa aca acc agg ttt ttc aac aga gat cct aat 1298 Leu Ser Ala Gly Thr Lys Thr Thr Arg Phe Phe Asn Arg Asp Pro Asn 420 425 430 ttg ggg tac cca ctg tgg agg ctt aag aca cct gaa gaa cat gaa ctg 1346 Leu Gly Tyr Pro Leu Trp Arg Leu Lys Thr Pro Glu Glu His Glu Leu 435 440 445 gag act gga ata aaa tca aaa gaa gct cgg aag tac att ttt aat tgc 1394 Glu Thr Gly Ile Lys Ser Lys Glu Ala Arg Lys Tyr Ile Phe Asn Cys 450 455 460 tta gat gac atg gct cag gtg aat atg tct aca gac ctg gag gga aca 1442 Leu Asp Asp Met Ala Gln Val Asn Met Ser Thr Asp Leu Glu Gly Thr 465 470 475 gac atg ttg gca gag aag gca gac cga aga gaa tac att gat ctg tta 1490 Asp Met Leu Ala Glu Lys Ala Asp Arg Arg Glu Tyr Ile Asp Leu Leu 480 485 490 495 aag aaa atg ctc aca att gat gca gat aag aga att acc cct cta aaa 1538 Lys Lys Met Leu Thr Ile Asp Ala Asp Lys Arg Ile Thr Pro Leu Lys 500 505 510 act ctt aac cat cag ttt gtg aca atg act cac ctt ttg gat ttt cca 1586 Thr Leu Asn His Gln Phe Val Thr Met Thr His Leu Leu Asp Phe Pro 515 520 525 cat agc aat cat gtt aag tct tgt ttt cag aac atg gag atc tgc aag 1634 His Ser Asn His Val Lys Ser Cys Phe Gln Asn Met Glu Ile Cys Lys 530 535 540 cgg agg gtt cac atg tat gat aca gtg agt cag atc aag agt ccc ttc 1682 Arg Arg Val His Met Tyr Asp Thr Val Ser Gln Ile Lys Ser Pro Phe 545 550 555 act aca cat gtt gcc cca aat aca agc aca aat cta acc atg agc ttc 1730 Thr Thr His Val Ala Pro Asn Thr Ser Thr Asn Leu Thr Met Ser Phe 560 565 570 575 agc aat cag ctc aat aca gtg cac aat cag gcc agt gtt cta gct tcc 1778 Ser Asn Gln Leu Asn Thr Val His Asn Gln Ala Ser Val Leu Ala Ser 580 585 590 agt tct act gca gca gct gct act ctt tct ctg gct aat tca gat gtc 1826 Ser Ser Thr Ala Ala Ala Ala Thr Leu Ser Leu Ala Asn Ser Asp Val 595 600 605 tca cta cta aac tac cag tca gct ttg tac cca tca tct gct gca cca 1874 Ser Leu Leu Asn Tyr Gln Ser Ala Leu Tyr Pro Ser Ser Ala Ala Pro 610 615 620 gtt cct gga gtt gcc cag cag ggt gtt tcc ttg cag cct gga acc acc 1922 Val Pro Gly Val Ala Gln Gln Gly Val Ser Leu Gln Pro Gly Thr Thr 625 630 635 cag att tgc act cag aca gat cca ttc caa cag aca ttt ata gta tgt 1970 Gln Ile Cys Thr Gln Thr Asp Pro Phe Gln Gln Thr Phe Ile Val Cys 640 645 650 655 cca cct gcg ttt caa act gga cta caa gca aca aca aag cat tct gga 2018 Pro Pro Ala Phe Gln Thr Gly Leu Gln Ala Thr Thr Lys His Ser Gly 660 665 670 ttc cct gtg agg atg gat aat gct gta ccg att gta ccc cag gca cca 2066 Phe Pro Val Arg Met Asp Asn Ala Val Pro Ile Val Pro Gln Ala Pro 675 680 685 gct gct cag cca cta cag att cag tca gga gtt ctc acg cag gga agc 2114 Ala Ala Gln Pro Leu Gln Ile Gln Ser Gly Val Leu Thr Gln Gly Ser 690 695 700 tgt aca cca cta atg gta gca act ctc cac cct caa gta gcc acc atc 2162 Cys Thr Pro Leu Met Val Ala Thr Leu His Pro Gln Val Ala Thr Ile 705 710 715 aca ccg cag tat gcg gtg ccc ttt act ctg agc tgc gca gcc ggc cgg 2210 Thr Pro Gln Tyr Ala Val Pro Phe Thr Leu Ser Cys Ala Ala Gly Arg 720 725 730 735 ccg gcg ctg gtt gaa cag act gcc gct gta ctg cag gcg tgg cct gga 2258 Pro Ala Leu Val Glu Gln Thr Ala Ala Val Leu Gln Ala Trp Pro Gly 740 745 750 ggg act cag caa att ctc ctg cct tca act tgg caa cag ttg cct ggg 2306 Gly Thr Gln Gln Ile Leu Leu Pro Ser Thr Trp Gln Gln Leu Pro Gly 755 760 765 gta gct cta cac aac tct gtc cag ccc aca gca atg att cca gag gcc 2354 Val Ala Leu His Asn Ser Val Gln Pro Thr Ala Met Ile Pro Glu Ala 770 775 780 atg ggg agt gga cag cag cta gct gac tgg agg aat gcc cac tct cat 2402 Met Gly Ser Gly Gln Gln Leu Ala Asp Trp Arg Asn Ala His Ser His 785 790 795 ggc aac cag tac agc act atc atg cag cag cca tcc ttg ctg act aac 2450 Gly Asn Gln Tyr Ser Thr Ile Met Gln Gln Pro Ser Leu Leu Thr Asn 800 805 810 815 cat gtg aca ttg gcc act gct cag cct ctg aat gtt ggt gtt gcc cat 2498 His Val Thr Leu Ala Thr Ala Gln Pro Leu Asn Val Gly Val Ala His 820 825 830 gtt gtc aga caa caa caa tcc agt tcc ctc cct tcg aag aag aat aag 2546 Val Val Arg Gln Gln Gln Ser Ser Ser Leu Pro Ser Lys Lys Asn Lys 835 840 845 cag tca gct cca gtc tct tcc aag tcc tct cta gat gtt ctg cct tcc 2594 Gln Ser Ala Pro Val Ser Ser Lys Ser Ser Leu Asp Val Leu Pro Ser 850 855 860 caa gtc tat tct ctg gtt ggg agc agt ccc ctc cgc acc aca tct tct 2642 Gln Val Tyr Ser Leu Val Gly Ser Ser Pro Leu Arg Thr Thr Ser Ser 865 870 875 tat aat tcc ttg gtc cct gtc caa gat cag cat cag ccc atc atc att 2690 Tyr Asn Ser Leu Val Pro Val Gln Asp Gln His Gln Pro Ile Ile Ile 880 885 890 895 cca gat act ccc agc cct cct gtg agt gtc atc act atc cga agt gac 2738 Pro Asp Thr Pro Ser Pro Pro Val Ser Val Ile Thr Ile Arg Ser Asp 900 905 910 act gat gag gaa gag gac aac aaa tac aag ccc agt agc tct gga ctg 2786 Thr Asp Glu Glu Glu Asp Asn Lys Tyr Lys Pro Ser Ser Ser Gly Leu 915 920 925 aag cca agg tct aat gtc atc agt tat gtc act gtc aat gat tct cca 2834 Lys Pro Arg Ser Asn Val Ile Ser Tyr Val Thr Val Asn Asp Ser Pro 930 935 940 gac tct gac tct tct ttg agc agc cct tat tcc act gat acc ctg agt 2882 Asp Ser Asp Ser Ser Leu Ser Ser Pro Tyr Ser Thr Asp Thr Leu Ser 945 950 955 gct ctc cga ggc aat agt gga tcc gtt ttg gag ggg cct ggc aga gtt 2930 Ala Leu Arg Gly Asn Ser Gly Ser Val Leu Glu Gly Pro Gly Arg Val 960 965 970 975 gtg gca gat ggc act ggc acc cgc act atc att gtg cct cca ctg aaa 2978 Val Ala Asp Gly Thr Gly Thr Arg Thr Ile Ile Val Pro Pro Leu Lys 980 985 990 act cag ctt ggt gac tgc act gta gca acc cag gcc tca ggt ctc ctg 3026 Thr Gln Leu Gly Asp Cys Thr Val Ala Thr Gln Ala Ser Gly Leu Leu 995 1000 1005 agc aat aag act aag cca gtc gct tca gtg agt ggg cag tca tct gga 3074 Ser Asn Lys Thr Lys Pro Val Ala Ser Val Ser Gly Gln Ser Ser Gly 1010 1015 1020 tgc tgt atc acc ccc aca ggg tat cga gct caa cgc ggg ggg acc agt 3122 Cys Cys Ile Thr Pro Thr Gly Tyr Arg Ala Gln Arg Gly Gly Thr Ser 1025 1030 1035 gca gca caa cca ctc aat ctt agc cag aac cag cag tca tcg gcg gct 3170 Ala Ala Gln Pro Leu Asn Leu Ser Gln Asn Gln Gln Ser Ser Ala Ala 1040 1045 1050 1055 cca acc tca cag gag aga agc agc aac cca gcc ccc cgc agg cag cag 3218 Pro Thr Ser Gln Glu Arg Ser Ser Asn Pro Ala Pro Arg Arg Gln Gln 1060 1065 1070 gcg ttt gtg gcc cct ctc tcc caa gcc ccc tac acc ttc cag cat ggc 3266 Ala Phe Val Ala Pro Leu Ser Gln Ala Pro Tyr Thr Phe Gln His Gly 1075 1080 1085 agc ccg cta cac tcg aca ggg cac cca cac ctt gcc ccg gcc cct gct 3314 Ser Pro Leu His Ser Thr Gly His Pro His Leu Ala Pro Ala Pro Ala 1090 1095 1100 cac ctg cca agc cag gct cat ctg tat acg tat gct gcc ccg act tct 3362 His Leu Pro Ser Gln Ala His Leu Tyr Thr Tyr Ala Ala Pro Thr Ser 1105 1110 1115 gct gct gca ctg ggc tca acc agc tcc att gct cat ctt ttc tcc cca 3410 Ala Ala Ala Leu Gly Ser Thr Ser Ser Ile Ala His Leu Phe Ser Pro 1120 1125 1130 1135 cag ggt tcc tca agg cat gct gca gcc tat acc act cac cct agc act 3458 Gln Gly Ser Ser Arg His Ala Ala Ala Tyr Thr Thr His Pro Ser Thr 1140 1145 1150 ttg gtg cac cag gtc cct gtc agt gtt ggg ccc agc ctc ctc act tct 3506 Leu Val His Gln Val Pro Val Ser Val Gly Pro Ser Leu Leu Thr Ser 1155 1160 1165 gcc agc gtg gcc cct gct cag tac caa cac cag ttt gcc acc caa tcc 3554 Ala Ser Val Ala Pro Ala Gln Tyr Gln His Gln Phe Ala Thr Gln Ser 1170 1175 1180 tac att ggg tct tcc cga ggc tca aca att tac act gga tac ccg ctg 3602 Tyr Ile Gly Ser Ser Arg Gly Ser Thr Ile Tyr Thr Gly Tyr Pro Leu 1185 1190 1195 agt cct acc aag atc agc cag tat tcc tac tta tag ttggtgagca 3648 Ser Pro Thr Lys Ile Ser Gln Tyr Ser Tyr Leu * 1200 1205 1210 tgagggagga ggaatcatgg ctaccttctc ctggccctgc gttcttaata ttgggctatg 3708 gagagatcct cctttaccct cttgaaattt cttagccagc aacttgttct gcaggggccc 3768 actgaagcag aaggtttttc tctgggggaa cctgtctcag tgttgactgc attgttgtag 3828 tcttcccaaa gtttgcccta tttttaaatt cattattttt gtgacagtaa ttttggtact 3888 tggaagagtt cagatgccca tcttctgcag ttaccaagga agagagattg ttctgaagtt 3948 accctctgaa aaatattttg tctctctgac ttgatttcta taaatgcttt taaaaacaag 4008 tgaagcccct ctttatttca ttttgtgtta ttgtgattgc tggtcaggaa aaatgctgat 4068 agaaggagtt gaaatctgat gacaaaaaaa gaaaaattac tttttgtttg tttataaact 4128 cagacttgcc tattttattt taaaagcggc ttacacaatc tcccttttgt ttattggaca 4188 tttaaactta cagagtttca gttttgtttt aatgtcatat tatacttaat gggcaattgt 4248 tatttttgca aaactggtta cgtattactc tgtgttacta ttgagattct ctcaattgct 4308 cctgtgtttg ttataaagta gtgtttaaaa ggcagctcac catttgctgg taacttaatg 4368 tgagagaatc catatctgcg tgaaaacacc aagtattctt tttaaatgaa gcaccatgaa 4428 ttctttttta aattattttt taaaagtctt tctctctctg attcagctta aattttttta 4488 tcgaaaaagc cattaaggtg gttattatta catggtggtg gtggttttat tatatgcaaa 4548 atctctgtct attatgagat actggcattg atgagctttg cctaaagatt agtatgaatt 4608 ttcagtaata cacctctgtt ttgctcatct ctcccttctg ttttatgtga tttgtttggg 4668 gagaaagcta aaaaaacctg aaaccagata agaacatttc ttgtgtatag cttttatact 4728 tcaaagtagc ttcctttgta tgccagcagc aaattgaatg ctctcttatt aagacttata 4788 taataagtgc atgtaggaat tgcaaaaaat attttaaaaa tttattactg aatttaaaaa 4848 tattttagaa gttttgtaat ggtggtgttt taatatttta cataattaaa tatgtacata 4908 ttgattagaa aaatataaca agcaattttt cctgctaacc caaaatgtta tttgtaatca 4968 aatgtgtagt gattacactt gaattgtgta cttagtgtgt atgtgatcct ccagtgttat 5028 cccggagatg gattgatgtc tccattgtat ttaaaccaaa atgaactgat acttgttgga 5088 atgtatgtga actaattgca attatattag agcatattac tgtagtgctg aatgagcagg 5148 ggcattgcct gcaaggagag gagacccttg gaattgtttt gcacaggtgt gtctggtgag 5208 gagtttttca gtgtgtgtct cttccttccc tttcttcctc cttcccttat tgagtgcctt 5268 atatgataat gtagtggtta atagagttta cagtgagctt gccttaggat ggaccagcaa 5328 gcccccgggg accctaagtt gttcaccggg atttatcaga acaggattag tagctggatt 5388 g 5389 89 1210 PRT Homo sapiens 89 Met Ala Ser Gln Leu Gln Val Phe Ser Pro Pro Ser Val Ser Ser Ser 1 5 10 15 Ala Phe Cys Ser Ala Lys Lys Leu Lys Ile Glu Pro Ser Gly Trp Asp 20 25 30 Val Ser Gly Gln Ser Ser Asn Asp Lys Tyr Tyr Thr His Ser Lys Thr 35 40 45 Leu Pro Ala Thr Gln Gly Gln Ala Asn Ser Ser His Gln Val Ala Asn 50 55 60 Phe Asn Ile Pro Ala Tyr Asp Gln Gly Leu Leu Leu Pro Ala Pro Ala 65 70 75 80 Val Glu His Ile Val Val Thr Ala Ala Asp Ser Ser Gly Ser Ala Ala 85 90 95 Thr Ser Thr Phe Gln Ser Ser Gln Thr Leu Thr Pro Arg Ser Asn Val 100 105 110 Ser Leu Leu Glu Pro Tyr Gln Lys Cys Gly Leu Lys Arg Lys Ser Glu 115 120 125 Glu Val Asp Ser Asn Gly Ser Val Gln Ile Ile Glu Glu His Pro Pro 130 135 140 Leu Met Leu Gln Asn Arg Thr Val Val Gly Ala Ala Ala Thr Thr Thr 145 150 155 160 Thr Val Thr Thr Lys Ser Ser Ser Ser Ser Gly Glu Gly Asp Tyr Gln 165 170 175 Leu Val Gln His Glu Ile Leu Cys Ser Met Thr Asn Ser Tyr Glu Val 180 185 190 Leu Glu Phe Leu Gly Arg Gly Thr Phe Gly Gln Val Ala Lys Cys Trp 195 200 205 Lys Arg Ser Thr Lys Glu Ile Val Ala Ile Lys Ile Leu Lys Asn His 210 215 220 Pro Ser Tyr Ala Arg Gln Gly Gln Ile Glu Val Ser Ile Leu Ser Arg 225 230 235 240 Leu Ser Ser Glu Asn Ala Asp Glu Tyr Asn Phe Val Arg Ser Tyr Glu 245 250 255 Cys Phe Gln His Lys Asn His Thr Cys Leu Val Phe Glu Met Leu Glu 260 265 270 Gln Asn Leu Tyr Asp Phe Leu Lys Gln Asn Lys Phe Ser Pro Leu Pro 275 280 285 Leu Lys Tyr Ile Arg Pro Ile Leu Gln Gln Val Ala Thr Ala Leu Met 290 295 300 Lys Leu Lys Ser Leu Gly Leu Ile His Ala Asp Leu Lys Pro Glu Asn 305 310 315 320 Ile Met Leu Val Asp Pro Val Arg Gln Pro Tyr Arg Val Lys Val Phe 325 330 335 Asp Phe Gly Ser Ala Ser His Val Ser Lys Ala Val Cys Ser Thr Tyr 340 345 350 Leu Gln Ser Arg Tyr Tyr Arg Ala Pro Glu Ile Ile Leu Gly Leu Pro 355 360 365 Phe Cys Glu Ala Ile Asp Met Trp Ser Leu Gly Cys Val Ile Ala Glu 370 375 380 Leu Phe Leu Gly Trp Pro Leu Tyr Pro Gly Ala Ser Glu Tyr Asp Gln 385 390 395 400 Ile Arg Tyr Ile Ser Gln Thr Gln Gly Leu Pro Ala Glu Tyr Leu Leu 405 410 415 Ser Ala Gly Thr Lys Thr Thr Arg Phe Phe Asn Arg Asp Pro Asn Leu 420 425 430 Gly Tyr Pro Leu Trp Arg Leu Lys Thr Pro Glu Glu His Glu Leu Glu 435 440 445 Thr Gly Ile Lys Ser Lys Glu Ala Arg Lys Tyr Ile Phe Asn Cys Leu 450 455 460 Asp Asp Met Ala Gln Val Asn Met Ser Thr Asp Leu Glu Gly Thr Asp 465 470 475 480 Met Leu Ala Glu Lys Ala Asp Arg Arg Glu Tyr Ile Asp Leu Leu Lys 485 490 495 Lys Met Leu Thr Ile Asp Ala Asp Lys Arg Ile Thr Pro Leu Lys Thr 500 505 510 Leu Asn His Gln Phe Val Thr Met Thr His Leu Leu Asp Phe Pro His 515 520 525 Ser Asn His Val Lys Ser Cys Phe Gln Asn Met Glu Ile Cys Lys Arg 530 535 540 Arg Val His Met Tyr Asp Thr Val Ser Gln Ile Lys Ser Pro Phe Thr 545 550 555 560 Thr His Val Ala Pro Asn Thr Ser Thr Asn Leu Thr Met Ser Phe Ser 565 570 575 Asn Gln Leu Asn Thr Val His Asn Gln Ala Ser Val Leu Ala Ser Ser 580 585 590 Ser Thr Ala Ala Ala Ala Thr Leu Ser Leu Ala Asn Ser Asp Val Ser 595 600 605 Leu Leu Asn Tyr Gln Ser Ala Leu Tyr Pro Ser Ser Ala Ala Pro Val 610 615 620 Pro Gly Val Ala Gln Gln Gly Val Ser Leu Gln Pro Gly Thr Thr Gln 625 630 635 640 Ile Cys Thr Gln Thr Asp Pro Phe Gln Gln Thr Phe Ile Val Cys Pro 645 650 655 Pro Ala Phe Gln Thr Gly Leu Gln Ala Thr Thr Lys His Ser Gly Phe 660 665 670 Pro Val Arg Met Asp Asn Ala Val Pro Ile Val Pro Gln Ala Pro Ala 675 680 685 Ala Gln Pro Leu Gln Ile Gln Ser Gly Val Leu Thr Gln Gly Ser Cys 690 695 700 Thr Pro Leu Met Val Ala Thr Leu His Pro Gln Val Ala Thr Ile Thr 705 710 715 720 Pro Gln Tyr Ala Val Pro Phe Thr Leu Ser Cys Ala Ala Gly Arg Pro 725 730 735 Ala Leu Val Glu Gln Thr Ala Ala Val Leu Gln Ala Trp Pro Gly Gly 740 745 750 Thr Gln Gln Ile Leu Leu Pro Ser Thr Trp Gln Gln Leu Pro Gly Val 755 760 765 Ala Leu His Asn Ser Val Gln Pro Thr Ala Met Ile Pro Glu Ala Met 770 775 780 Gly Ser Gly Gln Gln Leu Ala Asp Trp Arg Asn Ala His Ser His Gly 785 790 795 800 Asn Gln Tyr Ser Thr Ile Met Gln Gln Pro Ser Leu Leu Thr Asn His 805 810 815 Val Thr Leu Ala Thr Ala Gln Pro Leu Asn Val Gly Val Ala His Val 820 825 830 Val Arg Gln Gln Gln Ser Ser Ser Leu Pro Ser Lys Lys Asn Lys Gln 835 840 845 Ser Ala Pro Val Ser Ser Lys Ser Ser Leu Asp Val Leu Pro Ser Gln 850 855 860 Val Tyr Ser Leu Val Gly Ser Ser Pro Leu Arg Thr Thr Ser Ser Tyr 865 870 875 880 Asn Ser Leu Val Pro Val Gln Asp Gln His Gln Pro Ile Ile Ile Pro 885 890 895 Asp Thr Pro Ser Pro Pro Val Ser Val Ile Thr Ile Arg Ser Asp Thr 900 905 910 Asp Glu Glu Glu Asp Asn Lys Tyr Lys Pro Ser Ser Ser Gly Leu Lys 915 920 925 Pro Arg Ser Asn Val Ile Ser Tyr Val Thr Val Asn Asp Ser Pro Asp 930 935 940 Ser Asp Ser Ser Leu Ser Ser Pro Tyr Ser Thr Asp Thr Leu Ser Ala 945 950 955 960 Leu Arg Gly Asn Ser Gly Ser Val Leu Glu Gly Pro Gly Arg Val Val 965 970 975 Ala Asp Gly Thr Gly Thr Arg Thr Ile Ile Val Pro Pro Leu Lys Thr 980 985 990 Gln Leu Gly Asp Cys Thr Val Ala Thr Gln Ala Ser Gly Leu Leu Ser 995 1000 1005 Asn Lys Thr Lys Pro Val Ala Ser Val Ser Gly Gln Ser Ser Gly Cys 1010 1015 1020 Cys Ile Thr Pro Thr Gly Tyr Arg Ala Gln Arg Gly Gly Thr Ser Ala 1025 1030 1035 1040 Ala Gln Pro Leu Asn Leu Ser Gln Asn Gln Gln Ser Ser Ala Ala Pro 1045 1050 1055 Thr Ser Gln Glu Arg Ser Ser Asn Pro Ala Pro Arg Arg Gln Gln Ala 1060 1065 1070 Phe Val Ala Pro Leu Ser Gln Ala Pro Tyr Thr Phe Gln His Gly Ser 1075 1080 1085 Pro Leu His Ser Thr Gly His Pro His Leu Ala Pro Ala Pro Ala His 1090 1095 1100 Leu Pro Ser Gln Ala His Leu Tyr Thr Tyr Ala Ala Pro Thr Ser Ala 1105 1110 1115 1120 Ala Ala Leu Gly Ser Thr Ser Ser Ile Ala His Leu Phe Ser Pro Gln 1125 1130 1135 Gly Ser Ser Arg His Ala Ala Ala Tyr Thr Thr His Pro Ser Thr Leu 1140 1145 1150 Val His Gln Val Pro Val Ser Val Gly Pro Ser Leu Leu Thr Ser Ala 1155 1160 1165 Ser Val Ala Pro Ala Gln Tyr Gln His Gln Phe Ala Thr Gln Ser Tyr 1170 1175 1180 Ile Gly Ser Ser Arg Gly Ser Thr Ile Tyr Thr Gly Tyr Pro Leu Ser 1185 1190 1195 1200 Pro Thr Lys Ile Ser Gln Tyr Ser Tyr Leu 1205 1210 90 3633 DNA Homo sapiens CDS (1)...(3633) 90 atg gca tca cag ctg caa gtg ttt tcg ccc cca tca gtg tcg tcg agt 48 Met Ala Ser Gln Leu Gln Val Phe Ser Pro Pro Ser Val Ser Ser Ser 1 5 10 15 gcc ttc tgc agt gcg aag aaa ctg aaa ata gag ccc tct ggc tgg gat 96 Ala Phe Cys Ser Ala Lys Lys Leu Lys Ile Glu Pro Ser Gly Trp Asp 20 25 30 gtt tca gga cag agt agc aac gac aaa tat tat acc cac agc aaa acc 144 Val Ser Gly Gln Ser Ser Asn Asp Lys Tyr Tyr Thr His Ser Lys Thr 35 40 45 ctc cca gcc aca caa ggg caa gcc aac tcc tct cac cag gta gca aat 192 Leu Pro Ala Thr Gln Gly Gln Ala Asn Ser Ser His Gln Val Ala Asn 50 55 60 ttc aac atc cct gct tac gac cag ggc ctc ctc ctc cca gct cct gca 240 Phe Asn Ile Pro Ala Tyr Asp Gln Gly Leu Leu Leu Pro Ala Pro Ala 65 70 75 80 gtg gag cat att gtt gta aca gcc gct gat agc tcg ggc agt gct gct 288 Val Glu His Ile Val Val Thr Ala Ala Asp Ser Ser Gly Ser Ala Ala 85 90 95 aca tca acc ttc caa agc agc cag acc ctg act ccc aga agc aac gtt 336 Thr Ser Thr Phe Gln Ser Ser Gln Thr Leu Thr Pro Arg Ser Asn Val 100 105 110 tct ttg ctt gag cca tat caa aaa tgt gga ttg aaa cga aaa agt gag 384 Ser Leu Leu Glu Pro Tyr Gln Lys Cys Gly Leu Lys Arg Lys Ser Glu 115 120 125 gaa gtt gac agc aac ggt agt gtg cag atc ata gaa gaa cat ccc cct 432 Glu Val Asp Ser Asn Gly Ser Val Gln Ile Ile Glu Glu His Pro Pro 130 135 140 ctc atg ctg caa aac agg act gtg gtg ggt gct gct gcc aca acc acc 480 Leu Met Leu Gln Asn Arg Thr Val Val Gly Ala Ala Ala Thr Thr Thr 145 150 155 160 act gtg acc aca aag agt agc agt tcc agc gga gaa ggg gat tac cag 528 Thr Val Thr Thr Lys Ser Ser Ser Ser Ser Gly Glu Gly Asp Tyr Gln 165 170 175 ctg gtc cag cat gag atc ctt tgc tct atg acc aat agc tat gaa gtc 576 Leu Val Gln His Glu Ile Leu Cys Ser Met Thr Asn Ser Tyr Glu Val 180 185 190 ttg gag ttc cta ggc cgg ggg aca ttt gga cag gtg gct aag tgc tgg 624 Leu Glu Phe Leu Gly Arg Gly Thr Phe Gly Gln Val Ala Lys Cys Trp 195 200 205 aag agg agc acc aag gaa att gtg gct att aaa atc ttg aag aac cac 672 Lys Arg Ser Thr Lys Glu Ile Val Ala Ile Lys Ile Leu Lys Asn His 210 215 220 ccc tcc tat gcc aga caa gga cag att gaa gtg agc atc ctt tcc cgc 720 Pro Ser Tyr Ala Arg Gln Gly Gln Ile Glu Val Ser Ile Leu Ser Arg 225 230 235 240 cta agc agt gaa aat gct gat gag tat aat ttt gtc cgt tca tac gag 768 Leu Ser Ser Glu Asn Ala Asp Glu Tyr Asn Phe Val Arg Ser Tyr Glu 245 250 255 tgc ttt cag cat aag aat cac acc tgc ctt gtt ttt gaa atg ttg gag 816 Cys Phe Gln His Lys Asn His Thr Cys Leu Val Phe Glu Met Leu Glu 260 265 270 cag aac tta tat gat ttt cta aag caa aac aaa ttt agc cca ctg cca 864 Gln Asn Leu Tyr Asp Phe Leu Lys Gln Asn Lys Phe Ser Pro Leu Pro 275 280 285 ctc aag tac atc aga cca atc ttg cag cag gtg gcc aca gcc ttg atg 912 Leu Lys Tyr Ile Arg Pro Ile Leu Gln Gln Val Ala Thr Ala Leu Met 290 295 300 aag ctc aag agt ctt ggt ctg atc cac gct gac ctt aag cct gaa aac 960 Lys Leu Lys Ser Leu Gly Leu Ile His Ala Asp Leu Lys Pro Glu Asn 305 310 315 320 atc atg ctg gtt gat cca gtt cgc cag ccc tac cga gtg aag gtc ttt 1008 Ile Met Leu Val Asp Pro Val Arg Gln Pro Tyr Arg Val Lys Val Phe 325 330 335 gac ttt ggt tct gct agt cac gtt tcc aaa gct gtg tgc tca acc tac 1056 Asp Phe Gly Ser Ala Ser His Val Ser Lys Ala Val Cys Ser Thr Tyr 340 345 350 tta cag tca cgt tac tac aga gct cct gaa att att ctt ggg tta cca 1104 Leu Gln Ser Arg Tyr Tyr Arg Ala Pro Glu Ile Ile Leu Gly Leu Pro 355 360 365 ttt tgt gaa gct att gat atg tgg tca ctg ggc tgt gtg ata gct gag 1152 Phe Cys Glu Ala Ile Asp Met Trp Ser Leu Gly Cys Val Ile Ala Glu 370 375 380 ctg ttc ctg gga tgg cct ctt tat cct ggt gct tca gaa tat gat cag 1200 Leu Phe Leu Gly Trp Pro Leu Tyr Pro Gly Ala Ser Glu Tyr Asp Gln 385 390 395 400 att cgt tat att tca caa aca caa ggc ttg cca gct gaa tat ctt ctc 1248 Ile Arg Tyr Ile Ser Gln Thr Gln Gly Leu Pro Ala Glu Tyr Leu Leu 405 410 415 agt gcc gga aca aaa aca acc agg ttt ttc aac aga gat cct aat ttg 1296 Ser Ala Gly Thr Lys Thr Thr Arg Phe Phe Asn Arg Asp Pro Asn Leu 420 425 430 ggg tac cca ctg tgg agg ctt aag aca cct gaa gaa cat gaa ctg gag 1344 Gly Tyr Pro Leu Trp Arg Leu Lys Thr Pro Glu Glu His Glu Leu Glu 435 440 445 act gga ata aaa tca aaa gaa gct cgg aag tac att ttt aat tgc tta 1392 Thr Gly Ile Lys Ser Lys Glu Ala Arg Lys Tyr Ile Phe Asn Cys Leu 450 455 460 gat gac atg gct cag gtg aat atg tct aca gac ctg gag gga aca gac 1440 Asp Asp Met Ala Gln Val Asn Met Ser Thr Asp Leu Glu Gly Thr Asp 465 470 475 480 atg ttg gca gag aag gca gac cga aga gaa tac att gat ctg tta aag 1488 Met Leu Ala Glu Lys Ala Asp Arg Arg Glu Tyr Ile Asp Leu Leu Lys 485 490 495 aaa atg ctc aca att gat gca gat aag aga att acc cct cta aaa act 1536 Lys Met Leu Thr Ile Asp Ala Asp Lys Arg Ile Thr Pro Leu Lys Thr 500 505 510 ctt aac cat cag ttt gtg aca atg act cac ctt ttg gat ttt cca cat 1584 Leu Asn His Gln Phe Val Thr Met Thr His Leu Leu Asp Phe Pro His 515 520 525 agc aat cat gtt aag tct tgt ttt cag aac atg gag atc tgc aag cgg 1632 Ser Asn His Val Lys Ser Cys Phe Gln Asn Met Glu Ile Cys Lys Arg 530 535 540 agg gtt cac atg tat gat aca gtg agt cag atc aag agt ccc ttc act 1680 Arg Val His Met Tyr Asp Thr Val Ser Gln Ile Lys Ser Pro Phe Thr 545 550 555 560 aca cat gtt gcc cca aat aca agc aca aat cta acc atg agc ttc agc 1728 Thr His Val Ala Pro Asn Thr Ser Thr Asn Leu Thr Met Ser Phe Ser 565 570 575 aat cag ctc aat aca gtg cac aat cag gcc agt gtt cta gct tcc agt 1776 Asn Gln Leu Asn Thr Val His Asn Gln Ala Ser Val Leu Ala Ser Ser 580 585 590 tct act gca gca gct gct act ctt tct ctg gct aat tca gat gtc tca 1824 Ser Thr Ala Ala Ala Ala Thr Leu Ser Leu Ala Asn Ser Asp Val Ser 595 600 605 cta cta aac tac cag tca gct ttg tac cca tca tct gct gca cca gtt 1872 Leu Leu Asn Tyr Gln Ser Ala Leu Tyr Pro Ser Ser Ala Ala Pro Val 610 615 620 cct gga gtt gcc cag cag ggt gtt tcc ttg cag cct gga acc acc cag 1920 Pro Gly Val Ala Gln Gln Gly Val Ser Leu Gln Pro Gly Thr Thr Gln 625 630 635 640 att tgc act cag aca gat cca ttc caa cag aca ttt ata gta tgt cca 1968 Ile Cys Thr Gln Thr Asp Pro Phe Gln Gln Thr Phe Ile Val Cys Pro 645 650 655 cct gcg ttt caa act gga cta caa gca aca aca aag cat tct gga ttc 2016 Pro Ala Phe Gln Thr Gly Leu Gln Ala Thr Thr Lys His Ser Gly Phe 660 665 670 cct gtg agg atg gat aat gct gta ccg att gta ccc cag gca cca gct 2064 Pro Val Arg Met Asp Asn Ala Val Pro Ile Val Pro Gln Ala Pro Ala 675 680 685 gct cag cca cta cag att cag tca gga gtt ctc acg cag gga agc tgt 2112 Ala Gln Pro Leu Gln Ile Gln Ser Gly Val Leu Thr Gln Gly Ser Cys 690 695 700 aca cca cta atg gta gca act ctc cac cct caa gta gcc acc atc aca 2160 Thr Pro Leu Met Val Ala Thr Leu His Pro Gln Val Ala Thr Ile Thr 705 710 715 720 ccg cag tat gcg gtg ccc ttt act ctg agc tgc gca gcc ggc cgg ccg 2208 Pro Gln Tyr Ala Val Pro Phe Thr Leu Ser Cys Ala Ala Gly Arg Pro 725 730 735 gcg ctg gtt gaa cag act gcc gct gta ctg cag gcg tgg cct gga ggg 2256 Ala Leu Val Glu Gln Thr Ala Ala Val Leu Gln Ala Trp Pro Gly Gly 740 745 750 act cag caa att ctc ctg cct tca act tgg caa cag ttg cct ggg gta 2304 Thr Gln Gln Ile Leu Leu Pro Ser Thr Trp Gln Gln Leu Pro Gly Val 755 760 765 gct cta cac aac tct gtc cag ccc aca gca atg att cca gag gcc atg 2352 Ala Leu His Asn Ser Val Gln Pro Thr Ala Met Ile Pro Glu Ala Met 770 775 780 ggg agt gga cag cag cta gct gac tgg agg aat gcc cac tct cat ggc 2400 Gly Ser Gly Gln Gln Leu Ala Asp Trp Arg Asn Ala His Ser His Gly 785 790 795 800 aac cag tac agc act atc atg cag cag cca tcc ttg ctg act aac cat 2448 Asn Gln Tyr Ser Thr Ile Met Gln Gln Pro Ser Leu Leu Thr Asn His 805 810 815 gtg aca ttg gcc act gct cag cct ctg aat gtt ggt gtt gcc cat gtt 2496 Val Thr Leu Ala Thr Ala Gln Pro Leu Asn Val Gly Val Ala His Val 820 825 830 gtc aga caa caa caa tcc agt tcc ctc cct tcg aag aag aat aag cag 2544 Val Arg Gln Gln Gln Ser Ser Ser Leu Pro Ser Lys Lys Asn Lys Gln 835 840 845 tca gct cca gtc tct tcc aag tcc tct cta gat gtt ctg cct tcc caa 2592 Ser Ala Pro Val Ser Ser Lys Ser Ser Leu Asp Val Leu Pro Ser Gln 850 855 860 gtc tat tct ctg gtt ggg agc agt ccc ctc cgc acc aca tct tct tat 2640 Val Tyr Ser Leu Val Gly Ser Ser Pro Leu Arg Thr Thr Ser Ser Tyr 865 870 875 880 aat tcc ttg gtc cct gtc caa gat cag cat cag ccc atc atc att cca 2688 Asn Ser Leu Val Pro Val Gln Asp Gln His Gln Pro Ile Ile Ile Pro 885 890 895 gat act ccc agc cct cct gtg agt gtc atc act atc cga agt gac act 2736 Asp Thr Pro Ser Pro Pro Val Ser Val Ile Thr Ile Arg Ser Asp Thr 900 905 910 gat gag gaa gag gac aac aaa tac aag ccc agt agc tct gga ctg aag 2784 Asp Glu Glu Glu Asp Asn Lys Tyr Lys Pro Ser Ser Ser Gly Leu Lys 915 920 925 cca agg tct aat gtc atc agt tat gtc act gtc aat gat tct cca gac 2832 Pro Arg Ser Asn Val Ile Ser Tyr Val Thr Val Asn Asp Ser Pro Asp 930 935 940 tct gac tct tct ttg agc agc cct tat tcc act gat acc ctg agt gct 2880 Ser Asp Ser Ser Leu Ser Ser Pro Tyr Ser Thr Asp Thr Leu Ser Ala 945 950 955 960 ctc cga ggc aat agt gga tcc gtt ttg gag ggg cct ggc aga gtt gtg 2928 Leu Arg Gly Asn Ser Gly Ser Val Leu Glu Gly Pro Gly Arg Val Val 965 970 975 gca gat ggc act ggc acc cgc act atc att gtg cct cca ctg aaa act 2976 Ala Asp Gly Thr Gly Thr Arg Thr Ile Ile Val Pro Pro Leu Lys Thr 980 985 990 cag ctt ggt gac tgc act gta gca acc cag gcc tca ggt ctc ctg agc 3024 Gln Leu Gly Asp Cys Thr Val Ala Thr Gln Ala Ser Gly Leu Leu Ser 995 1000 1005 aat aag act aag cca gtc gct tca gtg agt ggg cag tca tct gga tgc 3072 Asn Lys Thr Lys Pro Val Ala Ser Val Ser Gly Gln Ser Ser Gly Cys 1010 1015 1020 tgt atc acc ccc aca ggg tat cga gct caa cgc ggg ggg acc agt gca 3120 Cys Ile Thr Pro Thr Gly Tyr Arg Ala Gln Arg Gly Gly Thr Ser Ala 1025 1030 1035 1040 gca caa cca ctc aat ctt agc cag aac cag cag tca tcg gcg gct cca 3168 Ala Gln Pro Leu Asn Leu Ser Gln Asn Gln Gln Ser Ser Ala Ala Pro 1045 1050 1055 acc tca cag gag aga agc agc aac cca gcc ccc cgc agg cag cag gcg 3216 Thr Ser Gln Glu Arg Ser Ser Asn Pro Ala Pro Arg Arg Gln Gln Ala 1060 1065 1070 ttt gtg gcc cct ctc tcc caa gcc ccc tac acc ttc cag cat ggc agc 3264 Phe Val Ala Pro Leu Ser Gln Ala Pro Tyr Thr Phe Gln His Gly Ser 1075 1080 1085 ccg cta cac tcg aca ggg cac cca cac ctt gcc ccg gcc cct gct cac 3312 Pro Leu His Ser Thr Gly His Pro His Leu Ala Pro Ala Pro Ala His 1090 1095 1100 ctg cca agc cag gct cat ctg tat acg tat gct gcc ccg act tct gct 3360 Leu Pro Ser Gln Ala His Leu Tyr Thr Tyr Ala Ala Pro Thr Ser Ala 1105 1110 1115 1120 gct gca ctg ggc tca acc agc tcc att gct cat ctt ttc tcc cca cag 3408 Ala Ala Leu Gly Ser Thr Ser Ser Ile Ala His Leu Phe Ser Pro Gln 1125 1130 1135 ggt tcc tca agg cat gct gca gcc tat acc act cac cct agc act ttg 3456 Gly Ser Ser Arg His Ala Ala Ala Tyr Thr Thr His Pro Ser Thr Leu 1140 1145 1150 gtg cac cag gtc cct gtc agt gtt ggg ccc agc ctc ctc act tct gcc 3504 Val His Gln Val Pro Val Ser Val Gly Pro Ser Leu Leu Thr Ser Ala 1155 1160 1165 agc gtg gcc cct gct cag tac caa cac cag ttt gcc acc caa tcc tac 3552 Ser Val Ala Pro Ala Gln Tyr Gln His Gln Phe Ala Thr Gln Ser Tyr 1170 1175 1180 att ggg tct tcc cga ggc tca aca att tac act gga tac ccg ctg agt 3600 Ile Gly Ser Ser Arg Gly Ser Thr Ile Tyr Thr Gly Tyr Pro Leu Ser 1185 1190 1195 1200 cct acc aag atc agc cag tat tcc tac tta tag 3633 Pro Thr Lys Ile Ser Gln Tyr Ser Tyr Leu * 1205 1210 91 270 PRT Artificial Sequence Ser/Thr kinase site consensus sequence 91 Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Gly Xaa Gly Xaa Xaa Xaa Xaa 1 5 10 15 Val Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Lys Xaa Xaa 20 25 30 Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 35 40 45 Xaa Glu Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 50 55 60 Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 65 70 75 80 Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 85 90 95 Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 100 105 110 Xaa Xaa Xaa Xaa His Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa His Arg Asp 115 120 125 Xaa Lys Xaa Xaa Asn Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 130 135 140 Xaa Xaa Xaa Xaa Xaa Xaa Lys Xaa Xaa Asp Phe Gly Xaa Xaa Xaa Xaa 145 150 155 160 Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 165 170 175 Xaa Pro Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Trp 180 185 190 Xaa Xaa Xaa Gly Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 195 200 205 Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 210 215 220 Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 225 230 235 240 Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Arg Xaa Xaa Xaa Xaa 245 250 255 Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa His Xaa Xaa Xaa 260 265 270 92 30 PRT Artificial Sequence ATP-binding region consensus sequence 92 Gly Xaa Gly Xaa Xaa Gly Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 1 5 10 15 Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Lys 20 25 30 93 214 PRT Artificial Sequence Amino acid consensus sequence 93 Tyr Glu Leu Leu Glu Lys Leu Gly Glu Gly Ser Phe Gly Lys Val Tyr 1 5 10 15 Lys Ala Lys His Lys Thr Gly Lys Ile Val Ala Val Lys Ile Leu Lys 20 25 30 Lys Glu Ser Leu Ser Leu Arg Glu Ile Gln Ile Leu Lys Arg Leu Ser 35 40 45 His Pro Asn Ile Val Arg Leu Leu Gly Val Phe Glu Asp Thr Asp Asp 50 55 60 His Leu Tyr Leu Val Met Glu Tyr Met Glu Gly Gly Asp Leu Phe Asp 65 70 75 80 Tyr Leu Arg Arg Asn Gly Pro Leu Ser Glu Lys Glu Ala Lys Lys Ile 85 90 95 Ala Leu Gln Ile Leu Arg Gly Leu Glu Tyr Leu His Ser Asn Gly Ile 100 105 110 Val His Arg Asp Leu Lys Pro Glu Asn Ile Leu Leu Asp Glu Asn Gly 115 120 125 Thr Val Lys Ile Ala Asp Phe Gly Leu Ala Arg Leu Leu Glu Lys Leu 130 135 140 Thr Thr Phe Val Gly Thr Pro Trp Tyr Met Met Ala Pro Glu Val Ile 145 150 155 160 Leu Glu Gly Arg Gly Tyr Ser Ser Lys Val Asp Val Trp Ser Leu Gly 165 170 175 Val Ile Leu Tyr Glu Leu Leu Thr Gly Gly Pro Leu Phe Pro Gly Ala 180 185 190 Asp Leu Pro Ala Phe Thr Gly Gly Asp Glu Val Asp Gln Leu Ile Ile 195 200 205 Phe Val Leu Lys Leu Pro 210 94 30 PRT Artificial Sequence Amino acid consensus sequence 94 Lys Asp Leu Leu Lys Lys Cys Leu Asn Lys Asp Pro Ser Lys Arg Pro 1 5 10 15 Gly Ser Ala Thr Ala Lys Glu Ile Leu Asn His Pro Trp Phe 20 25 30 95 158 PRT Artificial Sequence Amino acid consensus sequence 95 Leu Asn Ala Gly Thr Lys Thr Thr Arg Phe Phe Asn Arg Val Lys Ser 1 5 10 15 Glu Ser Pro Asn Asp Thr Asp Met Gly His Ser Tyr Trp Arg Leu Lys 20 25 30 Thr Pro Glu Glu His Glu Ala Glu Thr Gly Thr Ala Lys Ser Lys Glu 35 40 45 Ala Arg Lys Tyr Ile Phe Asn Cys Leu Asp Asp Ile Ala His Val Asn 50 55 60 Met Thr Met Asp Leu Glu Gly Ser Asp Met Leu Cys Glu Lys Ala Asp 65 70 75 80 Arg Arg Glu Phe Val Asp Leu Leu Lys Lys Met Leu Thr Ile Asp Ala 85 90 95 Asp Phe Arg Ile Thr Pro Ile Glu Thr Leu Asn His Pro Phe Val Thr 100 105 110 Met Thr His Leu Leu Asp Phe Pro His Ser Asn His Val Lys Ser Cys 115 120 125 Phe His Asn Met Glu Ile Cys Lys Lys Pro Gly Asn Ser Cys Asp Thr 130 135 140 Pro Asn His Ser Lys Thr Asn Leu Leu Thr Pro Val Ala Pro 145 150 155 96 135 PRT Artificial Sequence Amino acid consensus sequence 96 Pro Thr Ser Tyr Ser Ile Arg Pro Glu Asn Ala Val Pro Phe Val Thr 1 5 10 15 Gln Ala Pro Ala Ala Gln Pro Leu Gln Ile Gln Pro Gly Val Leu Ala 20 25 30 Gln Gln Ala Trp Pro Gly Gly Thr Gln Gln Ile Leu Leu Pro Pro Ala 35 40 45 Trp Gln Gln Leu Thr Gly Val Ala Pro His Thr Ser Val Gln Pro Ala 50 55 60 Ala Val Ile Pro Glu Ala Met Ala Gly Ser Gln Gln Leu Ala Asp Trp 65 70 75 80 Arg Asn Met His Ser His Gly Asn His Tyr Asn Thr Ile Met Gln Gln 85 90 95 Pro Ser Leu Leu Thr Asn His Val Thr Leu Ser Ala Ala Gln Pro Leu 100 105 110 Asn Val Gly Val Ala His Val Val Trp Gln Gln Pro Ser Ser Thr Lys 115 120 125 Pro Ser Lys Lys Cys Lys Gln 130 135 97 162 PRT Artificial Sequence Amino acid consensus sequence 97 Thr Gln Gln Ile Leu Leu Pro Pro Ala Trp Gln Gln Leu Thr Gly Val 1 5 10 15 Ala Pro His Thr Ser Val Gln Pro Ala Ala Val Ile Pro Glu Ala Met 20 25 30 Ala Gly Ser Gln Gln Leu Ala Asp Trp Arg Asn Met His Ser His Gly 35 40 45 Asn His Tyr Asn Thr Ile Met Gln Gln Pro Ser Leu Leu Thr Asn His 50 55 60 Val Thr Leu Ser Ala Ala Gln Pro Leu Asn Val Gly Val Ala His Val 65 70 75 80 Val Trp Gln Gln Pro Ser Ser Thr Lys Pro Ser Lys Lys Cys Lys Gln 85 90 95 His Gln Ile Leu Val Lys Leu Met Glu Trp Glu Pro Gly Arg Glu Glu 100 105 110 Ile Asn Ala Phe Ser Pro Val Asn Ser Leu Ser Asn Cys Glu Val Pro 115 120 125 His Ser Gln Phe Ile Ser Pro Pro Ile Ile Ser Gly Lys Glu Val Glu 130 135 140 Glu Ser Ser Pro Ile Arg Thr Thr Asp Asn His Asn Ser Pro Gly Pro 145 150 155 160 Cys Gln 98 55 PRT Artificial Sequence Amino acid consensus sequence 98 Ser Ile Arg Pro Glu Asn Ala Val Pro Phe Val Thr Gln Ala Pro Ala 1 5 10 15 Ala Gln Pro Leu Gln Ile Gln Pro Gly Val Leu Ala Gln Gln Ala Trp 20 25 30 Pro Gly Gly Thr Gln Gln Ile Leu Leu Pro Pro Ala Trp Gln Gln Leu 35 40 45 Thr Gly Val Ala Pro His Thr 50 55 99 188 PRT Artificial Sequence Amino acid consensus sequence 99 Gly Tyr Arg Gln Gln Arg Pro Gly Pro His Phe Gln Gln Gln Gln Pro 1 5 10 15 Leu Asn Leu Ser Gln Ala Gln His His Gly Ser Ala His Gln Glu Trp 20 25 30 Asn His Ser Ser Asn Phe Gly His Arg Arg Gln Gln Ala Tyr Ile Pro 35 40 45 Pro Thr Met Thr Gln Ala Pro Tyr Thr Phe Pro His Gly Ser Pro Asn 50 55 60 His Ser Thr Val His Pro His Leu Ala Gly Ala Pro Ala His Leu Pro 65 70 75 80 Gly Gln Pro His Leu Tyr Thr Tyr Pro Ala Pro Thr Ser Ala Ala Ala 85 90 95 Leu Gly Ser Thr Gly Pro Val Ala His Leu Leu Ala Ser Gln Gly Ser 100 105 110 Ser Arg His Met Val Gln His Thr Thr Tyr Asn Ile Ser His Pro Ser 115 120 125 Gly Ile Val His Gln Val Pro Val Ser Met Gly Pro Arg Leu Leu Pro 130 135 140 Ser Pro Thr Ile His Pro Thr Gln Tyr Lys Pro Gln Phe Ala Pro Gln 145 150 155 160 Ser Tyr Ile Ala Ala Ser Pro Ala Ser Thr Val Tyr Thr Gly Tyr Pro 165 170 175 Leu Ser Pro Thr Lys Ile Ser Gln Tyr Pro Tyr Met 180 185 100 1281 DNA Homo sapiens CDS (49)...(1239) 100 ctgcccggat agtataaatc gaggatccag gtctgggcag attcaacc atg gga gcc 57 Met Gly Ala 1 aac act tca aga aaa cca cca gtg ttt gat gaa aat gaa gat gtc aac 105 Asn Thr Ser Arg Lys Pro Pro Val Phe Asp Glu Asn Glu Asp Val Asn 5 10 15 ttt gac cac ttt gaa att ttg cga gcc att ggg aaa ggc agt ttt ggg 153 Phe Asp His Phe Glu Ile Leu Arg Ala Ile Gly Lys Gly Ser Phe Gly 20 25 30 35 aag gtc tgc att gta cag aag aat gat acc aag aag atg tac gca atg 201 Lys Val Cys Ile Val Gln Lys Asn Asp Thr Lys Lys Met Tyr Ala Met 40 45 50 aag tac atg aat aaa caa aag tgc gtg gag cgc aat gaa gtg aga aat 249 Lys Tyr Met Asn Lys Gln Lys Cys Val Glu Arg Asn Glu Val Arg Asn 55 60 65 gtc ttc aag gaa ctc cag atc atg cag ggt ctg gag cac cct ttc ctg 297 Val Phe Lys Glu Leu Gln Ile Met Gln Gly Leu Glu His Pro Phe Leu 70 75 80 gtt aat ttg tgg tat tcc ttc caa gat gag gaa gac atg ttc atg gtg 345 Val Asn Leu Trp Tyr Ser Phe Gln Asp Glu Glu Asp Met Phe Met Val 85 90 95 gtg gac ctc ctg ctg ggt gga gac ctg cgt tat cac ctg caa cag aac 393 Val Asp Leu Leu Leu Gly Gly Asp Leu Arg Tyr His Leu Gln Gln Asn 100 105 110 115 gtc cac ttc aag gaa gaa aca gtg aag ctc ttc atc tgt gag ctg gtc 441 Val His Phe Lys Glu Glu Thr Val Lys Leu Phe Ile Cys Glu Leu Val 120 125 130 atg gcc ctg gac tac ctg cag aac cag cgc atc att cac agg gat atg 489 Met Ala Leu Asp Tyr Leu Gln Asn Gln Arg Ile Ile His Arg Asp Met 135 140 145 aag cct gac aat att tta ctt gac gaa cat ggg cac gtg cac atc aca 537 Lys Pro Asp Asn Ile Leu Leu Asp Glu His Gly His Val His Ile Thr 150 155 160 gat ttc aac att gct gcg atg ctg ccc agg gag ata cag att acc acc 585 Asp Phe Asn Ile Ala Ala Met Leu Pro Arg Glu Ile Gln Ile Thr Thr 165 170 175 atg gct ggc acc aag cct tac atg gca cct gag atg ttc agc tcc aga 633 Met Ala Gly Thr Lys Pro Tyr Met Ala Pro Glu Met Phe Ser Ser Arg 180 185 190 195 aaa gga gca ggc tat tcc ttt gct gtt gac tgg tgg tcc ctg gga gtg 681 Lys Gly Ala Gly Tyr Ser Phe Ala Val Asp Trp Trp Ser Leu Gly Val 200 205 210 acg gca tat gaa ctg ctg aga ggc cgg aga ccg tat cat att cgc tcc 729 Thr Ala Tyr Glu Leu Leu Arg Gly Arg Arg Pro Tyr His Ile Arg Ser 215 220 225 agt act tcc agc aag gaa att gta cac acg ttt gag acg act gtt gta 777 Ser Thr Ser Ser Lys Glu Ile Val His Thr Phe Glu Thr Thr Val Val 230 235 240 act tac cct tct gcc tgg tca cag gaa atg gtg tca ctt ctt aaa aag 825 Thr Tyr Pro Ser Ala Trp Ser Gln Glu Met Val Ser Leu Leu Lys Lys 245 250 255 cta ctc gaa cct aat cca gac caa cga ttt tct cag tta tct gat gtc 873 Leu Leu Glu Pro Asn Pro Asp Gln Arg Phe Ser Gln Leu Ser Asp Val 260 265 270 275 cag aac ttc ccg tat atg aat gat ata aac tgg gat gca gtt ttt cag 921 Gln Asn Phe Pro Tyr Met Asn Asp Ile Asn Trp Asp Ala Val Phe Gln 280 285 290 aag agg ctc att cca ggt ttc att cct aat aaa ggc agg ctg aat tgt 969 Lys Arg Leu Ile Pro Gly Phe Ile Pro Asn Lys Gly Arg Leu Asn Cys 295 300 305 gat cct acc ttt gaa ctt gag gaa atg att ttg gag tcc aaa cct cta 1017 Asp Pro Thr Phe Glu Leu Glu Glu Met Ile Leu Glu Ser Lys Pro Leu 310 315 320 cat aag aaa aaa aag cgt ctg gca aag aag gag aag gat atg agg aaa 1065 His Lys Lys Lys Lys Arg Leu Ala Lys Lys Glu Lys Asp Met Arg Lys 325 330 335 tgc gat tct tct cag aca tgt ctt ctt caa gag cac ctt gac tct gtc 1113 Cys Asp Ser Ser Gln Thr Cys Leu Leu Gln Glu His Leu Asp Ser Val 340 345 350 355 cag aag gag ttc ata att ttc aac aga gaa aaa gta aac agg gac ttt 1161 Gln Lys Glu Phe Ile Ile Phe Asn Arg Glu Lys Val Asn Arg Asp Phe 360 365 370 aac aaa aga caa cca aat cta gcc ttg gaa caa acc aaa gac cca caa 1209 Asn Lys Arg Gln Pro Asn Leu Ala Leu Glu Gln Thr Lys Asp Pro Gln 375 380 385 ggt gag gat ggt cag aat aac aac ttg taa aggcctcatg tcttcttctt 1259 Gly Glu Asp Gly Gln Asn Asn Asn Leu * 390 395 gggacaatct catgccagaa ac 1281 101 396 PRT Homo sapiens 101 Met Gly Ala Asn Thr Ser Arg Lys Pro Pro Val Phe Asp Glu Asn Glu 1 5 10 15 Asp Val Asn Phe Asp His Phe Glu Ile Leu Arg Ala Ile Gly Lys Gly 20 25 30 Ser Phe Gly Lys Val Cys Ile Val Gln Lys Asn Asp Thr Lys Lys Met 35 40 45 Tyr Ala Met Lys Tyr Met Asn Lys Gln Lys Cys Val Glu Arg Asn Glu 50 55 60 Val Arg Asn Val Phe Lys Glu Leu Gln Ile Met Gln Gly Leu Glu His 65 70 75 80 Pro Phe Leu Val Asn Leu Trp Tyr Ser Phe Gln Asp Glu Glu Asp Met 85 90 95 Phe Met Val Val Asp Leu Leu Leu Gly Gly Asp Leu Arg Tyr His Leu 100 105 110 Gln Gln Asn Val His Phe Lys Glu Glu Thr Val Lys Leu Phe Ile Cys 115 120 125 Glu Leu Val Met Ala Leu Asp Tyr Leu Gln Asn Gln Arg Ile Ile His 130 135 140 Arg Asp Met Lys Pro Asp Asn Ile Leu Leu Asp Glu His Gly His Val 145 150 155 160 His Ile Thr Asp Phe Asn Ile Ala Ala Met Leu Pro Arg Glu Ile Gln 165 170 175 Ile Thr Thr Met Ala Gly Thr Lys Pro Tyr Met Ala Pro Glu Met Phe 180 185 190 Ser Ser Arg Lys Gly Ala Gly Tyr Ser Phe Ala Val Asp Trp Trp Ser 195 200 205 Leu Gly Val Thr Ala Tyr Glu Leu Leu Arg Gly Arg Arg Pro Tyr His 210 215 220 Ile Arg Ser Ser Thr Ser Ser Lys Glu Ile Val His Thr Phe Glu Thr 225 230 235 240 Thr Val Val Thr Tyr Pro Ser Ala Trp Ser Gln Glu Met Val Ser Leu 245 250 255 Leu Lys Lys Leu Leu Glu Pro Asn Pro Asp Gln Arg Phe Ser Gln Leu 260 265 270 Ser Asp Val Gln Asn Phe Pro Tyr Met Asn Asp Ile Asn Trp Asp Ala 275 280 285 Val Phe Gln Lys Arg Leu Ile Pro Gly Phe Ile Pro Asn Lys Gly Arg 290 295 300 Leu Asn Cys Asp Pro Thr Phe Glu Leu Glu Glu Met Ile Leu Glu Ser 305 310 315 320 Lys Pro Leu His Lys Lys Lys Lys Arg Leu Ala Lys Lys Glu Lys Asp 325 330 335 Met Arg Lys Cys Asp Ser Ser Gln Thr Cys Leu Leu Gln Glu His Leu 340 345 350 Asp Ser Val Gln Lys Glu Phe Ile Ile Phe Asn Arg Glu Lys Val Asn 355 360 365 Arg Asp Phe Asn Lys Arg Gln Pro Asn Leu Ala Leu Glu Gln Thr Lys 370 375 380 Asp Pro Gln Gly Glu Asp Gly Gln Asn Asn Asn Leu 385 390 395 102 1191 DNA Homo sapiens CDS (1)...(1191) 102 atg gga gcc aac act tca aga aaa cca cca gtg ttt gat gaa aat gaa 48 Met Gly Ala Asn Thr Ser Arg Lys Pro Pro Val Phe Asp Glu Asn Glu 1 5 10 15 gat gtc aac ttt gac cac ttt gaa att ttg cga gcc att ggg aaa ggc 96 Asp Val Asn Phe Asp His Phe Glu Ile Leu Arg Ala Ile Gly Lys Gly 20 25 30 agt ttt ggg aag gtc tgc att gta cag aag aat gat acc aag aag atg 144 Ser Phe Gly Lys Val Cys Ile Val Gln Lys Asn Asp Thr Lys Lys Met 35 40 45 tac gca atg aag tac atg aat aaa caa aag tgc gtg gag cgc aat gaa 192 Tyr Ala Met Lys Tyr Met Asn Lys Gln Lys Cys Val Glu Arg Asn Glu 50 55 60 gtg aga aat gtc ttc aag gaa ctc cag atc atg cag ggt ctg gag cac 240 Val Arg Asn Val Phe Lys Glu Leu Gln Ile Met Gln Gly Leu Glu His 65 70 75 80 cct ttc ctg gtt aat ttg tgg tat tcc ttc caa gat gag gaa gac atg 288 Pro Phe Leu Val Asn Leu Trp Tyr Ser Phe Gln Asp Glu Glu Asp Met 85 90 95 ttc atg gtg gtg gac ctc ctg ctg ggt gga gac ctg cgt tat cac ctg 336 Phe Met Val Val Asp Leu Leu Leu Gly Gly Asp Leu Arg Tyr His Leu 100 105 110 caa cag aac gtc cac ttc aag gaa gaa aca gtg aag ctc ttc atc tgt 384 Gln Gln Asn Val His Phe Lys Glu Glu Thr Val Lys Leu Phe Ile Cys 115 120 125 gag ctg gtc atg gcc ctg gac tac ctg cag aac cag cgc atc att cac 432 Glu Leu Val Met Ala Leu Asp Tyr Leu Gln Asn Gln Arg Ile Ile His 130 135 140 agg gat atg aag cct gac aat att tta ctt gac gaa cat ggg cac gtg 480 Arg Asp Met Lys Pro Asp Asn Ile Leu Leu Asp Glu His Gly His Val 145 150 155 160 cac atc aca gat ttc aac att gct gcg atg ctg ccc agg gag ata cag 528 His Ile Thr Asp Phe Asn Ile Ala Ala Met Leu Pro Arg Glu Ile Gln 165 170 175 att acc acc atg gct ggc acc aag cct tac atg gca cct gag atg ttc 576 Ile Thr Thr Met Ala Gly Thr Lys Pro Tyr Met Ala Pro Glu Met Phe 180 185 190 agc tcc aga aaa gga gca ggc tat tcc ttt gct gtt gac tgg tgg tcc 624 Ser Ser Arg Lys Gly Ala Gly Tyr Ser Phe Ala Val Asp Trp Trp Ser 195 200 205 ctg gga gtg acg gca tat gaa ctg ctg aga ggc cgg aga ccg tat cat 672 Leu Gly Val Thr Ala Tyr Glu Leu Leu Arg Gly Arg Arg Pro Tyr His 210 215 220 att cgc tcc agt act tcc agc aag gaa att gta cac acg ttt gag acg 720 Ile Arg Ser Ser Thr Ser Ser Lys Glu Ile Val His Thr Phe Glu Thr 225 230 235 240 act gtt gta act tac cct tct gcc tgg tca cag gaa atg gtg tca ctt 768 Thr Val Val Thr Tyr Pro Ser Ala Trp Ser Gln Glu Met Val Ser Leu 245 250 255 ctt aaa aag cta ctc gaa cct aat cca gac caa cga ttt tct cag tta 816 Leu Lys Lys Leu Leu Glu Pro Asn Pro Asp Gln Arg Phe Ser Gln Leu 260 265 270 tct gat gtc cag aac ttc ccg tat atg aat gat ata aac tgg gat gca 864 Ser Asp Val Gln Asn Phe Pro Tyr Met Asn Asp Ile Asn Trp Asp Ala 275 280 285 gtt ttt cag aag agg ctc att cca ggt ttc att cct aat aaa ggc agg 912 Val Phe Gln Lys Arg Leu Ile Pro Gly Phe Ile Pro Asn Lys Gly Arg 290 295 300 ctg aat tgt gat cct acc ttt gaa ctt gag gaa atg att ttg gag tcc 960 Leu Asn Cys Asp Pro Thr Phe Glu Leu Glu Glu Met Ile Leu Glu Ser 305 310 315 320 aaa cct cta cat aag aaa aaa aag cgt ctg gca aag aag gag aag gat 1008 Lys Pro Leu His Lys Lys Lys Lys Arg Leu Ala Lys Lys Glu Lys Asp 325 330 335 atg agg aaa tgc gat tct tct cag aca tgt ctt ctt caa gag cac ctt 1056 Met Arg Lys Cys Asp Ser Ser Gln Thr Cys Leu Leu Gln Glu His Leu 340 345 350 gac tct gtc cag aag gag ttc ata att ttc aac aga gaa aaa gta aac 1104 Asp Ser Val Gln Lys Glu Phe Ile Ile Phe Asn Arg Glu Lys Val Asn 355 360 365 agg gac ttt aac aaa aga caa cca aat cta gcc ttg gaa caa acc aaa 1152 Arg Asp Phe Asn Lys Arg Gln Pro Asn Leu Ala Leu Glu Gln Thr Lys 370 375 380 gac cca caa ggt gag gat ggt cag aat aac aac ttg taa 1191 Asp Pro Gln Gly Glu Asp Gly Gln Asn Asn Asn Leu * 385 390 395 103 270 PRT Artificial Sequence Ser/Thr kinase site consensus sequence 103 Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Gly Xaa Gly Xaa Xaa Xaa Xaa 1 5 10 15 Val Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Lys Xaa Xaa 20 25 30 Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 35 40 45 Xaa Glu Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 50 55 60 Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 65 70 75 80 Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 85 90 95 Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 100 105 110 Xaa Xaa Xaa Xaa His Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa His Arg Asp 115 120 125 Xaa Lys Xaa Xaa Asn Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 130 135 140 Xaa Xaa Xaa Xaa Xaa Xaa Lys Xaa Xaa Asp Phe Gly Xaa Xaa Xaa Xaa 145 150 155 160 Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 165 170 175 Xaa Pro Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Trp 180 185 190 Xaa Xaa Xaa Gly Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 195 200 205 Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 210 215 220 Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 225 230 235 240 Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Arg Xaa Xaa Xaa Xaa 245 250 255 Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa His Xaa Xaa Xaa 260 265 270 104 30 PRT Artificial Sequence ATP-binding region consensus sequence 104 Gly Xaa Gly Xaa Xaa Gly Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 1 5 10 15 Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Lys 20 25 30 105 278 PRT Artificial Sequence Eukaryotic protein kinase domain 105 Tyr Glu Leu Leu Glu Lys Leu Gly Glu Gly Ser Phe Gly Lys Val Tyr 1 5 10 15 Lys Ala Lys His Lys Thr Gly Lys Ile Val Ala Val Lys Ile Leu Lys 20 25 30 Lys Glu Ser Leu Ser Leu Arg Glu Ile Gln Ile Leu Lys Arg Leu Ser 35 40 45 His Pro Asn Ile Val Arg Leu Leu Gly Val Phe Glu Asp Thr Asp Asp 50 55 60 His Leu Tyr Leu Val Met Glu Tyr Met Glu Gly Gly Asp Leu Phe Asp 65 70 75 80 Tyr Leu Arg Arg Asn Gly Pro Leu Ser Glu Lys Glu Ala Lys Lys Ile 85 90 95 Ala Leu Gln Ile Leu Arg Gly Leu Glu Tyr Leu His Ser Asn Gly Ile 100 105 110 Val His Arg Asp Leu Lys Pro Glu Asn Ile Leu Leu Asp Glu Asn Gly 115 120 125 Thr Val Lys Ile Ala Asp Phe Gly Leu Ala Arg Leu Leu Glu Lys Leu 130 135 140 Thr Thr Phe Val Gly Thr Pro Trp Tyr Met Met Ala Pro Glu Val Ile 145 150 155 160 Leu Glu Gly Arg Gly Tyr Ser Ser Lys Val Asp Val Trp Ser Leu Gly 165 170 175 Val Ile Leu Tyr Glu Leu Leu Thr Gly Gly Pro Leu Phe Pro Gly Ala 180 185 190 Asp Leu Pro Ala Phe Thr Gly Gly Asp Glu Val Asp Gln Leu Ile Ile 195 200 205 Phe Val Leu Lys Leu Pro Phe Ser Asp Glu Leu Pro Lys Thr Arg Ile 210 215 220 Asp Pro Leu Glu Glu Leu Phe Arg Ile Lys Lys Arg Arg Leu Pro Leu 225 230 235 240 Pro Ser Asn Cys Ser Glu Glu Leu Lys Asp Leu Leu Lys Lys Cys Leu 245 250 255 Asn Lys Asp Pro Ser Lys Arg Pro Gly Ser Ala Thr Ala Lys Glu Ile 260 265 270 Leu Asn His Pro Trp Phe 275 106 20 PRT Artificial Sequence Protein kinase C terminal domain 106 Arg Glu Ile Asp Trp Asp Lys Leu Glu Asn Lys Glu Ile Glu Pro Pro 1 5 10 15 Phe Lys Pro Lys 20 107 129 PRT Artificial Sequence Kinase protein transferase ATP-binding serine/threonine-protein phosphorylation receptor tyrosine-protein precursor transmembrane 107 Pro Tyr Tyr Val Ser Met Lys Ser Met Ala Pro Glu Tyr Met Ala Pro 1 5 10 15 Glu Ser Ser Ala Thr Asn Tyr Gln Lys Tyr Ser Thr Lys Ser Asp Val 20 25 30 Trp Ser Phe Gly Val Ile Leu Tyr Glu Met Leu Thr Gly Lys Pro Pro 35 40 45 Phe Phe Pro Gly Glu Ser Glu Val Ser Glu Glu Glu Pro Tyr Gln Ser 50 55 60 Met Lys Asn Met Glu Val Leu Glu Met Gly Pro Glu Glu Thr Ile Gln 65 70 75 80 Lys Val Met Ser Lys Ile Val Glu Lys Lys Gly Glu Arg Met Pro Gln 85 90 95 Pro Ser Ser Ser Asn Cys Pro Glu Val Ser Gln Glu Ala Lys Asp Leu 100 105 110 Leu Lys Lys Cys Leu Gln Lys Asp Pro Glu Lys Arg Arg Pro Thr Phe 115 120 125 Glu 108 43 PRT Artificial Sequence Kinase protein transferase ATP-binding serine/threonine-protein phosphorylation receptor tyrosine-protein precursor transmembrane 108 Tyr Val His Gln Ile Ala Lys Gly Leu Glu Tyr Leu His Ser Lys Asn 1 5 10 15 Gln Lys His Gln Gly Ile Ile His Arg Ala Lys Lys Val Asp Leu Lys 20 25 30 Pro Glu Asn Ile Leu Leu Asp Glu Glu Ser His 35 40 109 53 PRT Artificial Sequence Kinase protein transferase ATP-binding serine/threonine-protein phosphorylation receptor tyrosine-protein precursor transmembrane 109 Tyr Glu Leu Leu Lys Lys Leu Leu Gly Lys Gly Ser Phe Gly Lys Val 1 5 10 15 Tyr Lys Ala Lys His Lys Ser Thr Ser Thr Thr Gly Glu Val Val Ala 20 25 30 Val Lys Val Met Lys Lys Lys Lys Val Met Glu Lys Ser Ser Lys Glu 35 40 45 Ser Ser Ser Lys Lys 50 110 31 PRT Artificial Sequence Kinase protein transferase ATP-binding serine/threonine-protein phosphorylation receptor tyrosine-protein precursor transmembrane 110 Ser Gln Glu Ala Lys Asp Leu Leu Lys Lys Cys Leu Gln Lys Asp Pro 1 5 10 15 Glu Lys Arg Arg Pro Thr Phe Glu Glu Ile Leu Gln His Pro Trp 20 25 30 111 87 PRT Artificial Sequence M03C11.1 protein 111 Tyr Thr Glu Arg Ile Asp Phe Lys Ser Val Phe Glu Lys Lys Pro Ser 1 5 10 15 Pro Val Phe Ile Pro Cys Lys Glu Gly Leu Asn Cys Asp Pro Met Tyr 20 25 30 Glu Leu Glu Glu Arg Ile Leu Val Ser Thr Pro Ile His Arg Arg Arg 35 40 45 Thr Asn His Asn Asn Ser Ser Gly Arg Ser Ser Ser Glu Pro Gln Asn 50 55 60 Ala Ala Leu Val Glu Val Ser Lys Ala Phe Ile Asp Phe Ser Arg His 65 70 75 80 Asn Val Lys Ile Glu Pro Asn 85 112 72 PRT Artificial Sequence F8K4.6 protein 112 Thr Asp Pro Ala Ile Trp Leu Lys Leu Glu Ala Ala Ile Glu Glu Phe 1 5 10 15 Ile Gln Ser Asn Pro Gln Val Phe Lys Asn Val Cys Glu Arg Leu Thr 20 25 30 Leu Pro Phe Leu Asn Asp Glu Lys Trp Cys Asp Asn Leu Lys Pro Arg 35 40 45 Phe Met Asn Gly Lys Leu Pro Asn Ser Arg Val Glu Ser Ser Pro Ser 50 55 60 Leu Gly Trp Arg Arg Asn Val Leu 65 70 113 3577 DNA Homo sapiens CDS (47)...(2386) 113 ctcctcagat cggtggacgt gctcgcctcc actcggggcc aggtct atg tcc cgg 55 Met Ser Arg 1 ttt ccc gca gtc gcg ggc agg gcg cca agg cgg cag gag gag ggt gag 103 Phe Pro Ala Val Ala Gly Arg Ala Pro Arg Arg Gln Glu Glu Gly Glu 5 10 15 cgg tca aga gac ctc cag gaa gag cgg ctc tcg gct gtt tgc atc gcc 151 Arg Ser Arg Asp Leu Gln Glu Glu Arg Leu Ser Ala Val Cys Ile Ala 20 25 30 35 gat aga gaa gag aaa gga tgc acg tcc cag gag gga gga act act cca 199 Asp Arg Glu Glu Lys Gly Cys Thr Ser Gln Glu Gly Gly Thr Thr Pro 40 45 50 act ttt cct att cag aaa caa aga aaa aag att att caa gct gtg agg 247 Thr Phe Pro Ile Gln Lys Gln Arg Lys Lys Ile Ile Gln Ala Val Arg 55 60 65 gac aat tca ttc ctt att gtt act gga aat aca gga agt ggt aaa aca 295 Asp Asn Ser Phe Leu Ile Val Thr Gly Asn Thr Gly Ser Gly Lys Thr 70 75 80 act caa ctc cca aaa tat cta tat gaa gca ggg ttt tca caa cat ggt 343 Thr Gln Leu Pro Lys Tyr Leu Tyr Glu Ala Gly Phe Ser Gln His Gly 85 90 95 atg att ggt gta act caa cca cga aaa gta gct gct ata tca gtt gct 391 Met Ile Gly Val Thr Gln Pro Arg Lys Val Ala Ala Ile Ser Val Ala 100 105 110 115 cag aga gta gct gaa gaa atg aaa tgc act ttg gga tcc aaa gta gga 439 Gln Arg Val Ala Glu Glu Met Lys Cys Thr Leu Gly Ser Lys Val Gly 120 125 130 tac caa gtt cgt ttt gat gat tgc agt tct aag gag aca gca atc aaa 487 Tyr Gln Val Arg Phe Asp Asp Cys Ser Ser Lys Glu Thr Ala Ile Lys 135 140 145 tat atg act gat gga tgt tta ctg aaa cat att ctg gga gac cca aat 535 Tyr Met Thr Asp Gly Cys Leu Leu Lys His Ile Leu Gly Asp Pro Asn 150 155 160 ctt acc aaa ttc agt gtc att att ttg gat gaa gcc cat gaa aga act 583 Leu Thr Lys Phe Ser Val Ile Ile Leu Asp Glu Ala His Glu Arg Thr 165 170 175 cta act aca gat atc tta ttt ggt tta ttg aag aag cta ttt cag gag 631 Leu Thr Thr Asp Ile Leu Phe Gly Leu Leu Lys Lys Leu Phe Gln Glu 180 185 190 195 aag tct cct aat agg aag gag cat tta aaa gtg gtg gta atg tca gca 679 Lys Ser Pro Asn Arg Lys Glu His Leu Lys Val Val Val Met Ser Ala 200 205 210 act atg gaa tta gcc aag ctc tct gca ttc ttt gga aat tgt cca ata 727 Thr Met Glu Leu Ala Lys Leu Ser Ala Phe Phe Gly Asn Cys Pro Ile 215 220 225 ttt gat ata cct gga agg ctt tat cca gtc aga gag aaa ttc tgc aat 775 Phe Asp Ile Pro Gly Arg Leu Tyr Pro Val Arg Glu Lys Phe Cys Asn 230 235 240 ttg att ggt cca cga gac aga gaa aat act gcg tat att caa gcg att 823 Leu Ile Gly Pro Arg Asp Arg Glu Asn Thr Ala Tyr Ile Gln Ala Ile 245 250 255 gtg aaa gtc acc atg gat atc cat ttg aat gaa atg gct gga gac atc 871 Val Lys Val Thr Met Asp Ile His Leu Asn Glu Met Ala Gly Asp Ile 260 265 270 275 ttg gtt ttt ctg act ggc cag ttt gaa ata gaa aaa agt tgt gag tta 919 Leu Val Phe Leu Thr Gly Gln Phe Glu Ile Glu Lys Ser Cys Glu Leu 280 285 290 ctt ttt cag atg gca gag tct gtt gat tat gat tat gat gtt caa gat 967 Leu Phe Gln Met Ala Glu Ser Val Asp Tyr Asp Tyr Asp Val Gln Asp 295 300 305 acc acc ctc gat ggc ttg tta ata ttg ccg tgt tat gga tca atg aca 1015 Thr Thr Leu Asp Gly Leu Leu Ile Leu Pro Cys Tyr Gly Ser Met Thr 310 315 320 aca gat caa cag agg agg ata ttt ttg cca cca cca cct gga att aga 1063 Thr Asp Gln Gln Arg Arg Ile Phe Leu Pro Pro Pro Pro Gly Ile Arg 325 330 335 aaa tgt gtc ata tcc acc aat att tct gca acg tct ttg aca ata gat 1111 Lys Cys Val Ile Ser Thr Asn Ile Ser Ala Thr Ser Leu Thr Ile Asp 340 345 350 355 gga atc aga tat gtg gta gat ggt ggc ttc gtg aag cag tta aat cac 1159 Gly Ile Arg Tyr Val Val Asp Gly Gly Phe Val Lys Gln Leu Asn His 360 365 370 aac ccc aga tta ggg ttg gac atc ctg gag gtg gtt cca att tca aag 1207 Asn Pro Arg Leu Gly Leu Asp Ile Leu Glu Val Val Pro Ile Ser Lys 375 380 385 agc gag gca tta cag cga agt ggc cga gct ggc agg act tct tca gga 1255 Ser Glu Ala Leu Gln Arg Ser Gly Arg Ala Gly Arg Thr Ser Ser Gly 390 395 400 aaa tgc ttt cgg atc tat agt aaa gat ttt tgg aac cag tgt atg cct 1303 Lys Cys Phe Arg Ile Tyr Ser Lys Asp Phe Trp Asn Gln Cys Met Pro 405 410 415 gac cat gtg atc cct gaa att aag aga act agt ttg aca tct gta gtt 1351 Asp His Val Ile Pro Glu Ile Lys Arg Thr Ser Leu Thr Ser Val Val 420 425 430 435 ctg acc tta aag tgc ctt gcc ata cac gat gtc ata agg ttt ccc tat 1399 Leu Thr Leu Lys Cys Leu Ala Ile His Asp Val Ile Arg Phe Pro Tyr 440 445 450 ttg gat cca cct aat gag aga ctt att tta gaa gct ctt aaa caa ctt 1447 Leu Asp Pro Pro Asn Glu Arg Leu Ile Leu Glu Ala Leu Lys Gln Leu 455 460 465 tac cag tgt gat gct att gac agg agt ggc cat gtc acc aga ttg ggt 1495 Tyr Gln Cys Asp Ala Ile Asp Arg Ser Gly His Val Thr Arg Leu Gly 470 475 480 ttg tct atg gtg gag ttt cct ttg cct cca cat ctg aca tgt gca gta 1543 Leu Ser Met Val Glu Phe Pro Leu Pro Pro His Leu Thr Cys Ala Val 485 490 495 ata aaa gct gct tcc ctg gat tgt gaa gat cta cta ctt cca ata gca 1591 Ile Lys Ala Ala Ser Leu Asp Cys Glu Asp Leu Leu Leu Pro Ile Ala 500 505 510 515 gca atg ttg tct gtg gaa aac gtc ttc att aga cct gtt gat cca gag 1639 Ala Met Leu Ser Val Glu Asn Val Phe Ile Arg Pro Val Asp Pro Glu 520 525 530 tac cag aag gaa gca gaa cag aga cat cga gaa ttg gca gct aaa gct 1687 Tyr Gln Lys Glu Ala Glu Gln Arg His Arg Glu Leu Ala Ala Lys Ala 535 540 545 gga gga ttt aat gac ttt gca act tta gct gtc atc ttt gaa caa tgc 1735 Gly Gly Phe Asn Asp Phe Ala Thr Leu Ala Val Ile Phe Glu Gln Cys 550 555 560 aaa tca agt gga gct cca gct tca tgg tgc caa aaa cac tgg att cat 1783 Lys Ser Ser Gly Ala Pro Ala Ser Trp Cys Gln Lys His Trp Ile His 565 570 575 tgg agg tgc tta ttt tct gca ttt cgt gtg gaa gct caa ctt cga gaa 1831 Trp Arg Cys Leu Phe Ser Ala Phe Arg Val Glu Ala Gln Leu Arg Glu 580 585 590 595 cta atc agg aag ctt aaa cag caa agt gat ttc cca aaa gag acc ttt 1879 Leu Ile Arg Lys Leu Lys Gln Gln Ser Asp Phe Pro Lys Glu Thr Phe 600 605 610 gaa ggc cct aaa cat gaa gta cta cga aga tgt ctt tgt gcg ggc tat 1927 Glu Gly Pro Lys His Glu Val Leu Arg Arg Cys Leu Cys Ala Gly Tyr 615 620 625 ttc aaa aat gta gct cga aga tct gtt ggg aga acg ttt tgc aca atg 1975 Phe Lys Asn Val Ala Arg Arg Ser Val Gly Arg Thr Phe Cys Thr Met 630 635 640 gat ggt cgt gga agc cca gtt cac att cat cct tcc tca gca ctt cat 2023 Asp Gly Arg Gly Ser Pro Val His Ile His Pro Ser Ser Ala Leu His 645 650 655 gaa cag gaa acc aaa ctt gaa tgg atc att ttt cat gag gta ttg gtt 2071 Glu Gln Glu Thr Lys Leu Glu Trp Ile Ile Phe His Glu Val Leu Val 660 665 670 675 acc acc aaa gtc tac gca aga att gta tgc cca atc cgt tat gaa tgg 2119 Thr Thr Lys Val Tyr Ala Arg Ile Val Cys Pro Ile Arg Tyr Glu Trp 680 685 690 gta aga gac ttg tta ccc aag ttg cat gaa ttt aat gca cat gat ttg 2167 Val Arg Asp Leu Leu Pro Lys Leu His Glu Phe Asn Ala His Asp Leu 695 700 705 agc agt gtg gcc cga cgt gaa gtg aga gaa gat gca aga agg aga tgg 2215 Ser Ser Val Ala Arg Arg Glu Val Arg Glu Asp Ala Arg Arg Arg Trp 710 715 720 aca aat aag gaa aat gta aag cag cta aag gat gga ata tcg aaa gac 2263 Thr Asn Lys Glu Asn Val Lys Gln Leu Lys Asp Gly Ile Ser Lys Asp 725 730 735 gtc tta aag aaa atg caa aga aga aat gat gac aaa tcc ata tct gat 2311 Val Leu Lys Lys Met Gln Arg Arg Asn Asp Asp Lys Ser Ile Ser Asp 740 745 750 755 gca cgg gct cgt ttc ctt gag aga aag cag cag agg acc cag gac cac 2359 Ala Arg Ala Arg Phe Leu Glu Arg Lys Gln Gln Arg Thr Gln Asp His 760 765 770 agt gac aca cga aag gaa aca ggc taa ggtggtgaac cctccaattc 2406 Ser Asp Thr Arg Lys Glu Thr Gly * 775 aggaagtggg aaaaggagcc aggaaatgtg cttctacttt gccagttatt tcagacagca 2466 ctaccaagag gaggtggtca gcacttgtta ttggcctatg aactaaaagc aaatcaaagc 2526 tcataaatca aagctcatca gttcccataa atgcagttgt caaagaaaag atttggttgc 2586 catagtcata agcaatgata catgaaacca atgaaagaca gtacatgtaa taatattttc 2646 ctcagtacaa ttttgctggc cttaactggt atcaaacgct gtcattgaga tgttttcaaa 2706 gaacattgag ttgtatttaa tcagcgtgta ctccatttgc attgaagcat taaaaattat 2766 ttttcttaaa atctctttaa ggccttcttg ttgctgttag aatagtgcta tatatcaggt 2826 atgtgaccat ttatttcaga aggctgaaca taagaggttt ctactcagca atacttagat 2886 gtctaactgt ttaattgcta cagagcttta tagatattta gagaaaagac ttaatcaatt 2946 agtaaataaa attgcctatg gcaggattct ttcttgaatt aatattaatc cttaaattga 3006 tttttctggg attatacaaa ttccttttta tataaaagta tattgtttaa aacagtagct 3066 atagccatta accaaaggac agatgatata tatatatatg atatatatat atatataagt 3126 tcttttttag ctgtacctac gtacttatat cagcaccatg tatgtaggtg tgatagtact 3186 ttcaaacagc gcctccacct ggcctactct gttatttcca cctgtttggg tagggccatt 3246 taacttccat tatgccaaac ttgggatggg attttcgaag cagacaacac tatttcatcg 3306 tgtttcaaat tggaaccttg aggctagtta gtatcacact caggccacac tcagcacttg 3366 cccactcttg tttactgcct tgtattctag ttatttgtgt atttgtctcc ctcactagat 3426 tatacgctcc ttgtgggcag ggactgtgtc ttttttcatc tttgtatctt tcatggacct 3486 agcatagtgc tttgcacata gtagtcactc agtgtttgtt aaataaagct attagtgtca 3546 ttaaaattca aaaaaaaaaa aaaaaaaaaa a 3577 114 779 PRT Homo sapiens 114 Met Ser Arg Phe Pro Ala Val Ala Gly Arg Ala Pro Arg Arg Gln Glu 1 5 10 15 Glu Gly Glu Arg Ser Arg Asp Leu Gln Glu Glu Arg Leu Ser Ala Val 20 25 30 Cys Ile Ala Asp Arg Glu Glu Lys Gly Cys Thr Ser Gln Glu Gly Gly 35 40 45 Thr Thr Pro Thr Phe Pro Ile Gln Lys Gln Arg Lys Lys Ile Ile Gln 50 55 60 Ala Val Arg Asp Asn Ser Phe Leu Ile Val Thr Gly Asn Thr Gly Ser 65 70 75 80 Gly Lys Thr Thr Gln Leu Pro Lys Tyr Leu Tyr Glu Ala Gly Phe Ser 85 90 95 Gln His Gly Met Ile Gly Val Thr Gln Pro Arg Lys Val Ala Ala Ile 100 105 110 Ser Val Ala Gln Arg Val Ala Glu Glu Met Lys Cys Thr Leu Gly Ser 115 120 125 Lys Val Gly Tyr Gln Val Arg Phe Asp Asp Cys Ser Ser Lys Glu Thr 130 135 140 Ala Ile Lys Tyr Met Thr Asp Gly Cys Leu Leu Lys His Ile Leu Gly 145 150 155 160 Asp Pro Asn Leu Thr Lys Phe Ser Val Ile Ile Leu Asp Glu Ala His 165 170 175 Glu Arg Thr Leu Thr Thr Asp Ile Leu Phe Gly Leu Leu Lys Lys Leu 180 185 190 Phe Gln Glu Lys Ser Pro Asn Arg Lys Glu His Leu Lys Val Val Val 195 200 205 Met Ser Ala Thr Met Glu Leu Ala Lys Leu Ser Ala Phe Phe Gly Asn 210 215 220 Cys Pro Ile Phe Asp Ile Pro Gly Arg Leu Tyr Pro Val Arg Glu Lys 225 230 235 240 Phe Cys Asn Leu Ile Gly Pro Arg Asp Arg Glu Asn Thr Ala Tyr Ile 245 250 255 Gln Ala Ile Val Lys Val Thr Met Asp Ile His Leu Asn Glu Met Ala 260 265 270 Gly Asp Ile Leu Val Phe Leu Thr Gly Gln Phe Glu Ile Glu Lys Ser 275 280 285 Cys Glu Leu Leu Phe Gln Met Ala Glu Ser Val Asp Tyr Asp Tyr Asp 290 295 300 Val Gln Asp Thr Thr Leu Asp Gly Leu Leu Ile Leu Pro Cys Tyr Gly 305 310 315 320 Ser Met Thr Thr Asp Gln Gln Arg Arg Ile Phe Leu Pro Pro Pro Pro 325 330 335 Gly Ile Arg Lys Cys Val Ile Ser Thr Asn Ile Ser Ala Thr Ser Leu 340 345 350 Thr Ile Asp Gly Ile Arg Tyr Val Val Asp Gly Gly Phe Val Lys Gln 355 360 365 Leu Asn His Asn Pro Arg Leu Gly Leu Asp Ile Leu Glu Val Val Pro 370 375 380 Ile Ser Lys Ser Glu Ala Leu Gln Arg Ser Gly Arg Ala Gly Arg Thr 385 390 395 400 Ser Ser Gly Lys Cys Phe Arg Ile Tyr Ser Lys Asp Phe Trp Asn Gln 405 410 415 Cys Met Pro Asp His Val Ile Pro Glu Ile Lys Arg Thr Ser Leu Thr 420 425 430 Ser Val Val Leu Thr Leu Lys Cys Leu Ala Ile His Asp Val Ile Arg 435 440 445 Phe Pro Tyr Leu Asp Pro Pro Asn Glu Arg Leu Ile Leu Glu Ala Leu 450 455 460 Lys Gln Leu Tyr Gln Cys Asp Ala Ile Asp Arg Ser Gly His Val Thr 465 470 475 480 Arg Leu Gly Leu Ser Met Val Glu Phe Pro Leu Pro Pro His Leu Thr 485 490 495 Cys Ala Val Ile Lys Ala Ala Ser Leu Asp Cys Glu Asp Leu Leu Leu 500 505 510 Pro Ile Ala Ala Met Leu Ser Val Glu Asn Val Phe Ile Arg Pro Val 515 520 525 Asp Pro Glu Tyr Gln Lys Glu Ala Glu Gln Arg His Arg Glu Leu Ala 530 535 540 Ala Lys Ala Gly Gly Phe Asn Asp Phe Ala Thr Leu Ala Val Ile Phe 545 550 555 560 Glu Gln Cys Lys Ser Ser Gly Ala Pro Ala Ser Trp Cys Gln Lys His 565 570 575 Trp Ile His Trp Arg Cys Leu Phe Ser Ala Phe Arg Val Glu Ala Gln 580 585 590 Leu Arg Glu Leu Ile Arg Lys Leu Lys Gln Gln Ser Asp Phe Pro Lys 595 600 605 Glu Thr Phe Glu Gly Pro Lys His Glu Val Leu Arg Arg Cys Leu Cys 610 615 620 Ala Gly Tyr Phe Lys Asn Val Ala Arg Arg Ser Val Gly Arg Thr Phe 625 630 635 640 Cys Thr Met Asp Gly Arg Gly Ser Pro Val His Ile His Pro Ser Ser 645 650 655 Ala Leu His Glu Gln Glu Thr Lys Leu Glu Trp Ile Ile Phe His Glu 660 665 670 Val Leu Val Thr Thr Lys Val Tyr Ala Arg Ile Val Cys Pro Ile Arg 675 680 685 Tyr Glu Trp Val Arg Asp Leu Leu Pro Lys Leu His Glu Phe Asn Ala 690 695 700 His Asp Leu Ser Ser Val Ala Arg Arg Glu Val Arg Glu Asp Ala Arg 705 710 715 720 Arg Arg Trp Thr Asn Lys Glu Asn Val Lys Gln Leu Lys Asp Gly Ile 725 730 735 Ser Lys Asp Val Leu Lys Lys Met Gln Arg Arg Asn Asp Asp Lys Ser 740 745 750 Ile Ser Asp Ala Arg Ala Arg Phe Leu Glu Arg Lys Gln Gln Arg Thr 755 760 765 Gln Asp His Ser Asp Thr Arg Lys Glu Thr Gly 770 775 115 2340 DNA Homo sapiens CDS (1)...(2340) 115 atg tcc cgg ttt ccc gca gtc gcg ggc agg gcg cca agg cgg cag gag 48 Met Ser Arg Phe Pro Ala Val Ala Gly Arg Ala Pro Arg Arg Gln Glu 1 5 10 15 gag ggt gag cgg tca aga gac ctc cag gaa gag cgg ctc tcg gct gtt 96 Glu Gly Glu Arg Ser Arg Asp Leu Gln Glu Glu Arg Leu Ser Ala Val 20 25 30 tgc atc gcc gat aga gaa gag aaa gga tgc acg tcc cag gag gga gga 144 Cys Ile Ala Asp Arg Glu Glu Lys Gly Cys Thr Ser Gln Glu Gly Gly 35 40 45 act act cca act ttt cct att cag aaa caa aga aaa aag att att caa 192 Thr Thr Pro Thr Phe Pro Ile Gln Lys Gln Arg Lys Lys Ile Ile Gln 50 55 60 gct gtg agg gac aat tca ttc ctt att gtt act gga aat aca gga agt 240 Ala Val Arg Asp Asn Ser Phe Leu Ile Val Thr Gly Asn Thr Gly Ser 65 70 75 80 ggt aaa aca act caa ctc cca aaa tat cta tat gaa gca ggg ttt tca 288 Gly Lys Thr Thr Gln Leu Pro Lys Tyr Leu Tyr Glu Ala Gly Phe Ser 85 90 95 caa cat ggt atg att ggt gta act caa cca cga aaa gta gct gct ata 336 Gln His Gly Met Ile Gly Val Thr Gln Pro Arg Lys Val Ala Ala Ile 100 105 110 tca gtt gct cag aga gta gct gaa gaa atg aaa tgc act ttg gga tcc 384 Ser Val Ala Gln Arg Val Ala Glu Glu Met Lys Cys Thr Leu Gly Ser 115 120 125 aaa gta gga tac caa gtt cgt ttt gat gat tgc agt tct aag gag aca 432 Lys Val Gly Tyr Gln Val Arg Phe Asp Asp Cys Ser Ser Lys Glu Thr 130 135 140 gca atc aaa tat atg act gat gga tgt tta ctg aaa cat att ctg gga 480 Ala Ile Lys Tyr Met Thr Asp Gly Cys Leu Leu Lys His Ile Leu Gly 145 150 155 160 gac cca aat ctt acc aaa ttc agt gtc att att ttg gat gaa gcc cat 528 Asp Pro Asn Leu Thr Lys Phe Ser Val Ile Ile Leu Asp Glu Ala His 165 170 175 gaa aga act cta act aca gat atc tta ttt ggt tta ttg aag aag cta 576 Glu Arg Thr Leu Thr Thr Asp Ile Leu Phe Gly Leu Leu Lys Lys Leu 180 185 190 ttt cag gag aag tct cct aat agg aag gag cat tta aaa gtg gtg gta 624 Phe Gln Glu Lys Ser Pro Asn Arg Lys Glu His Leu Lys Val Val Val 195 200 205 atg tca gca act atg gaa tta gcc aag ctc tct gca ttc ttt gga aat 672 Met Ser Ala Thr Met Glu Leu Ala Lys Leu Ser Ala Phe Phe Gly Asn 210 215 220 tgt cca ata ttt gat ata cct gga agg ctt tat cca gtc aga gag aaa 720 Cys Pro Ile Phe Asp Ile Pro Gly Arg Leu Tyr Pro Val Arg Glu Lys 225 230 235 240 ttc tgc aat ttg att ggt cca cga gac aga gaa aat act gcg tat att 768 Phe Cys Asn Leu Ile Gly Pro Arg Asp Arg Glu Asn Thr Ala Tyr Ile 245 250 255 caa gcg att gtg aaa gtc acc atg gat atc cat ttg aat gaa atg gct 816 Gln Ala Ile Val Lys Val Thr Met Asp Ile His Leu Asn Glu Met Ala 260 265 270 gga gac atc ttg gtt ttt ctg act ggc cag ttt gaa ata gaa aaa agt 864 Gly Asp Ile Leu Val Phe Leu Thr Gly Gln Phe Glu Ile Glu Lys Ser 275 280 285 tgt gag tta ctt ttt cag atg gca gag tct gtt gat tat gat tat gat 912 Cys Glu Leu Leu Phe Gln Met Ala Glu Ser Val Asp Tyr Asp Tyr Asp 290 295 300 gtt caa gat acc acc ctc gat ggc ttg tta ata ttg ccg tgt tat gga 960 Val Gln Asp Thr Thr Leu Asp Gly Leu Leu Ile Leu Pro Cys Tyr Gly 305 310 315 320 tca atg aca aca gat caa cag agg agg ata ttt ttg cca cca cca cct 1008 Ser Met Thr Thr Asp Gln Gln Arg Arg Ile Phe Leu Pro Pro Pro Pro 325 330 335 gga att aga aaa tgt gtc ata tcc acc aat att tct gca acg tct ttg 1056 Gly Ile Arg Lys Cys Val Ile Ser Thr Asn Ile Ser Ala Thr Ser Leu 340 345 350 aca ata gat gga atc aga tat gtg gta gat ggt ggc ttc gtg aag cag 1104 Thr Ile Asp Gly Ile Arg Tyr Val Val Asp Gly Gly Phe Val Lys Gln 355 360 365 tta aat cac aac ccc aga tta ggg ttg gac atc ctg gag gtg gtt cca 1152 Leu Asn His Asn Pro Arg Leu Gly Leu Asp Ile Leu Glu Val Val Pro 370 375 380 att tca aag agc gag gca tta cag cga agt ggc cga gct ggc agg act 1200 Ile Ser Lys Ser Glu Ala Leu Gln Arg Ser Gly Arg Ala Gly Arg Thr 385 390 395 400 tct tca gga aaa tgc ttt cgg atc tat agt aaa gat ttt tgg aac cag 1248 Ser Ser Gly Lys Cys Phe Arg Ile Tyr Ser Lys Asp Phe Trp Asn Gln 405 410 415 tgt atg cct gac cat gtg atc cct gaa att aag aga act agt ttg aca 1296 Cys Met Pro Asp His Val Ile Pro Glu Ile Lys Arg Thr Ser Leu Thr 420 425 430 tct gta gtt ctg acc tta aag tgc ctt gcc ata cac gat gtc ata agg 1344 Ser Val Val Leu Thr Leu Lys Cys Leu Ala Ile His Asp Val Ile Arg 435 440 445 ttt ccc tat ttg gat cca cct aat gag aga ctt att tta gaa gct ctt 1392 Phe Pro Tyr Leu Asp Pro Pro Asn Glu Arg Leu Ile Leu Glu Ala Leu 450 455 460 aaa caa ctt tac cag tgt gat gct att gac agg agt ggc cat gtc acc 1440 Lys Gln Leu Tyr Gln Cys Asp Ala Ile Asp Arg Ser Gly His Val Thr 465 470 475 480 aga ttg ggt ttg tct atg gtg gag ttt cct ttg cct cca cat ctg aca 1488 Arg Leu Gly Leu Ser Met Val Glu Phe Pro Leu Pro Pro His Leu Thr 485 490 495 tgt gca gta ata aaa gct gct tcc ctg gat tgt gaa gat cta cta ctt 1536 Cys Ala Val Ile Lys Ala Ala Ser Leu Asp Cys Glu Asp Leu Leu Leu 500 505 510 cca ata gca gca atg ttg tct gtg gaa aac gtc ttc att aga cct gtt 1584 Pro Ile Ala Ala Met Leu Ser Val Glu Asn Val Phe Ile Arg Pro Val 515 520 525 gat cca gag tac cag aag gaa gca gaa cag aga cat cga gaa ttg gca 1632 Asp Pro Glu Tyr Gln Lys Glu Ala Glu Gln Arg His Arg Glu Leu Ala 530 535 540 gct aaa gct gga gga ttt aat gac ttt gca act tta gct gtc atc ttt 1680 Ala Lys Ala Gly Gly Phe Asn Asp Phe Ala Thr Leu Ala Val Ile Phe 545 550 555 560 gaa caa tgc aaa tca agt gga gct cca gct tca tgg tgc caa aaa cac 1728 Glu Gln Cys Lys Ser Ser Gly Ala Pro Ala Ser Trp Cys Gln Lys His 565 570 575 tgg att cat tgg agg tgc tta ttt tct gca ttt cgt gtg gaa gct caa 1776 Trp Ile His Trp Arg Cys Leu Phe Ser Ala Phe Arg Val Glu Ala Gln 580 585 590 ctt cga gaa cta atc agg aag ctt aaa cag caa agt gat ttc cca aaa 1824 Leu Arg Glu Leu Ile Arg Lys Leu Lys Gln Gln Ser Asp Phe Pro Lys 595 600 605 gag acc ttt gaa ggc cct aaa cat gaa gta cta cga aga tgt ctt tgt 1872 Glu Thr Phe Glu Gly Pro Lys His Glu Val Leu Arg Arg Cys Leu Cys 610 615 620 gcg ggc tat ttc aaa aat gta gct cga aga tct gtt ggg aga acg ttt 1920 Ala Gly Tyr Phe Lys Asn Val Ala Arg Arg Ser Val Gly Arg Thr Phe 625 630 635 640 tgc aca atg gat ggt cgt gga agc cca gtt cac att cat cct tcc tca 1968 Cys Thr Met Asp Gly Arg Gly Ser Pro Val His Ile His Pro Ser Ser 645 650 655 gca ctt cat gaa cag gaa acc aaa ctt gaa tgg atc att ttt cat gag 2016 Ala Leu His Glu Gln Glu Thr Lys Leu Glu Trp Ile Ile Phe His Glu 660 665 670 gta ttg gtt acc acc aaa gtc tac gca aga att gta tgc cca atc cgt 2064 Val Leu Val Thr Thr Lys Val Tyr Ala Arg Ile Val Cys Pro Ile Arg 675 680 685 tat gaa tgg gta aga gac ttg tta ccc aag ttg cat gaa ttt aat gca 2112 Tyr Glu Trp Val Arg Asp Leu Leu Pro Lys Leu His Glu Phe Asn Ala 690 695 700 cat gat ttg agc agt gtg gcc cga cgt gaa gtg aga gaa gat gca aga 2160 His Asp Leu Ser Ser Val Ala Arg Arg Glu Val Arg Glu Asp Ala Arg 705 710 715 720 agg aga tgg aca aat aag gaa aat gta aag cag cta aag gat gga ata 2208 Arg Arg Trp Thr Asn Lys Glu Asn Val Lys Gln Leu Lys Asp Gly Ile 725 730 735 tcg aaa gac gtc tta aag aaa atg caa aga aga aat gat gac aaa tcc 2256 Ser Lys Asp Val Leu Lys Lys Met Gln Arg Arg Asn Asp Asp Lys Ser 740 745 750 ata tct gat gca cgg gct cgt ttc ctt gag aga aag cag cag agg acc 2304 Ile Ser Asp Ala Arg Ala Arg Phe Leu Glu Arg Lys Gln Gln Arg Thr 755 760 765 cag gac cac agt gac aca cga aag gaa aca ggc taa 2340 Gln Asp His Ser Asp Thr Arg Lys Glu Thr Gly * 770 775 116 86 PRT Artificial Sequence Amino acid consensus sequence 116 Glu Glu Leu Lys Lys Leu Ile Leu Val Ala Thr Pro Gly Arg Leu Leu 1 5 10 15 Asp His Leu Glu Asn Gly Ser Leu Leu Glu Lys Arg Leu Lys Leu Lys 20 25 30 Asn Leu Lys Leu Leu Val Leu Asp Glu Ala Asp Arg Met Leu Asp Met 35 40 45 Gly Lys Ala His Gly Phe Gly Pro Asp Leu Glu Glu Gln Thr Leu Leu 50 55 60 Phe Ser Ala Thr Leu Pro Glu Val Glu Arg Leu Ala Lys Leu Phe Leu 65 70 75 80 Leu Arg Ile Lys Gln Lys 85 117 87 PRT Artificial Sequence Amino acid consensus sequence 117 Asp Glu Leu Ala Lys Phe Leu Lys Glu Leu Phe Pro Lys Leu Pro Gly 1 5 10 15 Ile Lys Val Ala Arg Leu His Gly Gly Leu Ser Gln Glu Glu Arg Glu 20 25 30 Glu Ile Leu Glu Lys Phe Arg Asn Gly Lys Ser Lys Val Leu Val Ala 35 40 45 Thr Asp Val Ala Ala Arg Gly Ile Asp Ile Pro Asp Val Asn Leu Val 50 55 60 Ile Asn Tyr Asp Leu Pro Trp Asn Pro Glu Ser Tyr Ile Gln Arg Ile 65 70 75 80 Gly Arg Ala Gly Arg Ala Gly 85 118 86 PRT Artificial Sequence Amino acid consensus sequence 118 Leu Asp Ala Glu Lys Phe Ser Glu Tyr Phe Gly Asn Cys Pro Ile Ile 1 5 10 15 Glu Val Pro Gly Arg Thr Tyr Pro Val Glu Val Tyr Tyr Thr Lys Glu 20 25 30 Thr Thr Glu Pro Glu Glu Asp Tyr Ile Glu Ala Ala Ile Arg Thr Val 35 40 45 Ile Gln Ile His Met Thr Glu Pro Ala Pro Gly Asp Ile Leu Val Phe 50 55 60 Leu Thr Gly Gln Glu Glu Ile Glu Glu Ala Cys Glu Arg Leu Lys Glu 65 70 75 80 Arg Met Lys Gln Leu Glu 85 119 745 PRT Homo sapiens 119 Arg Glu Arg Glu Lys Glu Lys Glu Lys Glu Leu Arg Ala Ser Thr Asn 1 5 10 15 Ala Met Leu Ile Ser Ala Gly Leu Pro Pro Leu Lys Ala Ser His Ser 20 25 30 Ala His Ser Thr His Ser Ala His Ser Thr His Ser Thr His Ser Ala 35 40 45 His Ser Thr His Ala Gly His Ala Gly His Thr Ser Leu Pro Gln Cys 50 55 60 Ile Asn Pro Phe Thr Asn Leu Pro His Thr Pro Arg Tyr Tyr Asp Ile 65 70 75 80 Leu Lys Lys Arg Leu Gln Leu Pro Val Trp Glu Tyr Lys Asp Arg Phe 85 90 95 Thr Asp Ile Leu Val Arg His Gln Ser Phe Val Leu Val Gly Glu Thr 100 105 110 Gly Ser Gly Lys Thr Thr Gln Ile Pro Gln Trp Cys Val Glu Tyr Met 115 120 125 Arg Ser Leu Pro Gly Pro Lys Arg Gly Val Ala Cys Thr Gln Pro Arg 130 135 140 Arg Val Ala Ala Met Ser Val Ala Gln Arg Val Ala Asp Glu Met Asp 145 150 155 160 Val Met Leu Gly Gln Glu Val Gly Tyr Ser Ile Arg Phe Glu Asp Cys 165 170 175 Ser Ser Ala Lys Thr Ile Leu Lys Tyr Met Thr Asp Gly Met Leu Leu 180 185 190 Arg Glu Ala Met Asn Asp Pro Leu Leu Glu Arg Tyr Gly Val Ile Ile 195 200 205 Leu Asp Glu Ala His Glu Arg Thr Leu Ala Thr Asp Ile Leu Met Gly 210 215 220 Val Leu Lys Glu Val Val Arg Gln Arg Ser Asp Leu Lys Val Ile Val 225 230 235 240 Met Ser Ala Thr Leu Asp Ala Gly Lys Phe Gln Ile Tyr Phe Asp Asn 245 250 255 Cys Pro Leu Leu Thr Ile Pro Gly Arg Thr His Pro Val Glu Ile Phe 260 265 270 Tyr Thr Pro Glu Pro Glu Arg Asp Tyr Leu Glu Ala Ala Ile Arg Thr 275 280 285 Val Ile Gln Ile His Met Cys Glu Glu Glu Glu Gly Asp Leu Leu Leu 290 295 300 Phe Leu Thr Gly Gln Glu Glu Ile Asp Glu Ala Cys Lys Arg Ile Lys 305 310 315 320 Arg Glu Val Asp Asp Leu Gly Pro Glu Val Gly Asp Ile Lys Ile Ile 325 330 335 Pro Leu Tyr Ser Thr Leu Pro Pro Gln Gln Gln Gln Arg Ile Phe Glu 340 345 350 Pro Pro Pro Pro Lys Lys Gln Asn Gly Ala Ile Gly Arg Lys Val Val 355 360 365 Val Ser Thr Asn Ile Ala Glu Thr Ser Leu Thr Ile Asp Gly Val Val 370 375 380 Phe Val Ile Asp Pro Gly Phe Ala Lys Gln Lys Val Tyr Asn Pro Arg 385 390 395 400 Ile Arg Val Glu Ser Leu Leu Val Thr Ala Ile Ser Lys Ala Ser Ala 405 410 415 Gln Gln Arg Ala Gly Arg Ala Gly Arg Thr Arg Pro Gly Lys Cys Phe 420 425 430 Arg Leu Tyr Thr Glu Lys Ala Tyr Lys Thr Glu Met Gln Asp Asn Thr 435 440 445 Tyr Pro Glu Ile Leu Arg Ser Asn Leu Gly Ser Val Val Leu Gln Leu 450 455 460 Lys Lys Leu Gly Ile Asp Asp Leu Val His Phe Asp Phe Met Asp Pro 465 470 475 480 Pro Ala Pro Glu Thr Leu Met Arg Ala Leu Glu Leu Leu Asn Tyr Leu 485 490 495 Ala Ala Leu Asn Asp Asp Gly Asp Leu Thr Glu Leu Gly Ser Met Met 500 505 510 Ala Glu Phe Pro Leu Asp Pro Gln Leu Ala Lys Met Val Ile Ala Ser 515 520 525 Cys Asp Tyr Asn Cys Ser Asn Glu Val Leu Ser Ile Thr Ala Met Leu 530 535 540 Ser Val Pro Gln Cys Phe Val Arg Pro Thr Glu Ala Lys Lys Ala Ala 545 550 555 560 Asp Glu Ala Lys Met Arg Phe Ala His Ile Asp Gly Asp His Leu Thr 565 570 575 Leu Leu Asn Val Tyr His Ala Phe Lys Gln Asn His Glu Ser Val Gln 580 585 590 Trp Cys Tyr Asp Asn Phe Ile Asn Tyr Arg Ser Leu Met Ser Ala Asp 595 600 605 Asn Val Arg Gln Gln Leu Ser Arg Ile Met Asp Arg Phe Asn Leu Pro 610 615 620 Arg Arg Ser Thr Asp Phe Thr Ser Arg Asp Tyr Tyr Ile Asn Ile Arg 625 630 635 640 Lys Ala Leu Val Thr Gly Tyr Phe Met Gln Val Ala His Leu Glu Arg 645 650 655 Thr Gly His Tyr Leu Thr Val Lys Asp Asn Gln Val Val Gln Leu His 660 665 670 Pro Ser Thr Val Leu Asp His Lys Pro Glu Trp Val Leu Tyr Asn Glu 675 680 685 Phe Val Leu Thr Thr Lys Asn Tyr Ile Arg Thr Cys Thr Asp Ile Lys 690 695 700 Pro Glu Trp Leu Val Lys Ile Ala Pro Gln Tyr Tyr Asp Met Ser Asn 705 710 715 720 Phe Pro Gln Cys Glu Ala Lys Arg Gln Leu Asp Arg Ile Ile Ala Lys 725 730 735 Leu Gln Ser Lys Glu Tyr Ser Gln Tyr 740 745 120 10 PRT Artificial Sequence DEAH-box subfamily ATP-dependent helicases signature 120 Xaa Xaa Xaa Xaa Xaa Asp Glu Xaa His Xaa 1 5 10 121 8 PRT Artificial Sequence ATP/GTP-binding motif ′A′ (P-loop) signature 121 Xaa Xaa Xaa Xaa Xaa Gly Lys Xaa 1 5 122 1649 DNA Homo sapiens CDS (213)...(1073) 122 gcgctgggtc cccgaggccc ggcccctccc cgggaggagg tgggcttcga gtcacgtgac 60 ccgtgcccta cgggaggggg tgcggtcggg gacccggcag gaggcggccg agaagagagg 120 accgtggggg cgttcgcgtg gctcccagcc cgggacccca cccccgctgg acagtggggg 180 aaactgaggc ctgagcgggc ccacacagga cc atg aag gtg ctt ctc ctc aca 233 Met Lys Val Leu Leu Leu Thr 1 5 ggg ctg ggg gcc ctg ttc ttc gcc tat tat tgg gat gac aac ttc gac 281 Gly Leu Gly Ala Leu Phe Phe Ala Tyr Tyr Trp Asp Asp Asn Phe Asp 10 15 20 cca gcc agc ctc cag gga gcg cga gtg ctg ctg aca ggg gcc aac gct 329 Pro Ala Ser Leu Gln Gly Ala Arg Val Leu Leu Thr Gly Ala Asn Ala 25 30 35 ggt gtt ggt gag gag ctg gcc tat cac tac gcg cgt ctg ggc tcc cac 377 Gly Val Gly Glu Glu Leu Ala Tyr His Tyr Ala Arg Leu Gly Ser His 40 45 50 55 ctg gtg ctc act gcc cac act gag gct ctc ctg cag aag gtg gta ggg 425 Leu Val Leu Thr Ala His Thr Glu Ala Leu Leu Gln Lys Val Val Gly 60 65 70 aac tgc cgg aag ctg ggc gcc ccc aag gtc ttc tac atc gcg gcg gac 473 Asn Cys Arg Lys Leu Gly Ala Pro Lys Val Phe Tyr Ile Ala Ala Asp 75 80 85 atg gcc tcc cct gag gcg ccc gag agc gtg gtg cag ttt gcg ctg gac 521 Met Ala Ser Pro Glu Ala Pro Glu Ser Val Val Gln Phe Ala Leu Asp 90 95 100 aag ctg ggc ggg ctg gac tac ctc gtg ctg aac cac atc ggc ggc gcc 569 Lys Leu Gly Gly Leu Asp Tyr Leu Val Leu Asn His Ile Gly Gly Ala 105 110 115 ccg gcc ggc acg cga gcc cgc agc ccc cag gca act cgc tgg ctc atg 617 Pro Ala Gly Thr Arg Ala Arg Ser Pro Gln Ala Thr Arg Trp Leu Met 120 125 130 135 cag gta aac ttt gtg agc tac gtg caa ctg acg tcg cgg gcg ctg ccc 665 Gln Val Asn Phe Val Ser Tyr Val Gln Leu Thr Ser Arg Ala Leu Pro 140 145 150 agc ctg acg gac agc aag ggc tcc ctg gtg gtg gtg tcc tcg ctg ctc 713 Ser Leu Thr Asp Ser Lys Gly Ser Leu Val Val Val Ser Ser Leu Leu 155 160 165 ggc cgc gtg ccc acg tcg ttc tcc act ccc tac tcg gcg gcc aag ttt 761 Gly Arg Val Pro Thr Ser Phe Ser Thr Pro Tyr Ser Ala Ala Lys Phe 170 175 180 gcg ctg gac ggc ttc ttc ggc tcc ctg cgg cgg gag ctg gac gtg cag 809 Ala Leu Asp Gly Phe Phe Gly Ser Leu Arg Arg Glu Leu Asp Val Gln 185 190 195 gac gtg aac gtg gcc atc acc atg tgc gtc ctg ggc ctc cga gat cgc 857 Asp Val Asn Val Ala Ile Thr Met Cys Val Leu Gly Leu Arg Asp Arg 200 205 210 215 gcc tcc gcc gcc gag gca gtc agg gga gtc acg agg gtc aag gcg gcc 905 Ala Ser Ala Ala Glu Ala Val Arg Gly Val Thr Arg Val Lys Ala Ala 220 225 230 ccg ggg ccc aag gca gcc ctg gcc gtg atc cgc ggc ggc gcc acg cgc 953 Pro Gly Pro Lys Ala Ala Leu Ala Val Ile Arg Gly Gly Ala Thr Arg 235 240 245 gcg gcc ggc gtc ttc tac ccg tgg cgt ttc cgc ctg ctg tgc ttg ctc 1001 Ala Ala Gly Val Phe Tyr Pro Trp Arg Phe Arg Leu Leu Cys Leu Leu 250 255 260 cgg cgc tgg cta ccg cgc ccg cgg gcc tgg ttt atc cgc cag gag ctc 1049 Arg Arg Trp Leu Pro Arg Pro Arg Ala Trp Phe Ile Arg Gln Glu Leu 265 270 275 aac gtc acg gcc gcg gca gcc tga gcaccggggg gtgcccctcc agtcccagac 1103 Asn Val Thr Ala Ala Ala Ala * 280 285 ggcaatgttc ctccctccaa ctgtccctgg agccagaaca ctcacagaga cacccctgag 1163 agggtggcca cagcccaaga tgaagtcatc aagacagaaa agcaaaaccg agaaaaacga 1223 cgggcacctg gaaccagtca cggcttggga ggtgcaggtg ccccgtgtta ggcgcctttg 1283 tcggggactt gcaaggcctc acctgtttgg ccatgattga tgacgtgact gcttccattt 1343 tgcagatgag gaaactaagg ctcagagagg ccacgccacc cttgagccac ccatggaccc 1403 ctctccatct cctgcctgcg cctttaagtc cctgatttat tctttccatt cattccatct 1463 gggaggaacc cccccaactc ctgccagctt cccctagctg gggtctctgg tactcttcac 1523 acctgcaggg gcgtctacac tgttcgtcta cctggtggca gggtctgagc gggaggagga 1583 gggaaagagt gtgttctgag ctggacccag cctcttgttc gagaataaaa actcttcttc 1643 tcttgc 1649 123 286 PRT Homo sapiens 123 Met Lys Val Leu Leu Leu Thr Gly Leu Gly Ala Leu Phe Phe Ala Tyr 1 5 10 15 Tyr Trp Asp Asp Asn Phe Asp Pro Ala Ser Leu Gln Gly Ala Arg Val 20 25 30 Leu Leu Thr Gly Ala Asn Ala Gly Val Gly Glu Glu Leu Ala Tyr His 35 40 45 Tyr Ala Arg Leu Gly Ser His Leu Val Leu Thr Ala His Thr Glu Ala 50 55 60 Leu Leu Gln Lys Val Val Gly Asn Cys Arg Lys Leu Gly Ala Pro Lys 65 70 75 80 Val Phe Tyr Ile Ala Ala Asp Met Ala Ser Pro Glu Ala Pro Glu Ser 85 90 95 Val Val Gln Phe Ala Leu Asp Lys Leu Gly Gly Leu Asp Tyr Leu Val 100 105 110 Leu Asn His Ile Gly Gly Ala Pro Ala Gly Thr Arg Ala Arg Ser Pro 115 120 125 Gln Ala Thr Arg Trp Leu Met Gln Val Asn Phe Val Ser Tyr Val Gln 130 135 140 Leu Thr Ser Arg Ala Leu Pro Ser Leu Thr Asp Ser Lys Gly Ser Leu 145 150 155 160 Val Val Val Ser Ser Leu Leu Gly Arg Val Pro Thr Ser Phe Ser Thr 165 170 175 Pro Tyr Ser Ala Ala Lys Phe Ala Leu Asp Gly Phe Phe Gly Ser Leu 180 185 190 Arg Arg Glu Leu Asp Val Gln Asp Val Asn Val Ala Ile Thr Met Cys 195 200 205 Val Leu Gly Leu Arg Asp Arg Ala Ser Ala Ala Glu Ala Val Arg Gly 210 215 220 Val Thr Arg Val Lys Ala Ala Pro Gly Pro Lys Ala Ala Leu Ala Val 225 230 235 240 Ile Arg Gly Gly Ala Thr Arg Ala Ala Gly Val Phe Tyr Pro Trp Arg 245 250 255 Phe Arg Leu Leu Cys Leu Leu Arg Arg Trp Leu Pro Arg Pro Arg Ala 260 265 270 Trp Phe Ile Arg Gln Glu Leu Asn Val Thr Ala Ala Ala Ala 275 280 285 124 861 DNA Homo sapiens CDS (1)...(861) 124 atg aag gtg ctt ctc ctc aca ggg ctg ggg gcc ctg ttc ttc gcc tat 48 Met Lys Val Leu Leu Leu Thr Gly Leu Gly Ala Leu Phe Phe Ala Tyr 1 5 10 15 tat tgg gat gac aac ttc gac cca gcc agc ctc cag gga gcg cga gtg 96 Tyr Trp Asp Asp Asn Phe Asp Pro Ala Ser Leu Gln Gly Ala Arg Val 20 25 30 ctg ctg aca ggg gcc aac gct ggt gtt ggt gag gag ctg gcc tat cac 144 Leu Leu Thr Gly Ala Asn Ala Gly Val Gly Glu Glu Leu Ala Tyr His 35 40 45 tac gcg cgt ctg ggc tcc cac ctg gtg ctc act gcc cac act gag gct 192 Tyr Ala Arg Leu Gly Ser His Leu Val Leu Thr Ala His Thr Glu Ala 50 55 60 ctc ctg cag aag gtg gta ggg aac tgc cgg aag ctg ggc gcc ccc aag 240 Leu Leu Gln Lys Val Val Gly Asn Cys Arg Lys Leu Gly Ala Pro Lys 65 70 75 80 gtc ttc tac atc gcg gcg gac atg gcc tcc cct gag gcg ccc gag agc 288 Val Phe Tyr Ile Ala Ala Asp Met Ala Ser Pro Glu Ala Pro Glu Ser 85 90 95 gtg gtg cag ttt gcg ctg gac aag ctg ggc ggg ctg gac tac ctc gtg 336 Val Val Gln Phe Ala Leu Asp Lys Leu Gly Gly Leu Asp Tyr Leu Val 100 105 110 ctg aac cac atc ggc ggc gcc ccg gcc ggc acg cga gcc cgc agc ccc 384 Leu Asn His Ile Gly Gly Ala Pro Ala Gly Thr Arg Ala Arg Ser Pro 115 120 125 cag gca act cgc tgg ctc atg cag gta aac ttt gtg agc tac gtg caa 432 Gln Ala Thr Arg Trp Leu Met Gln Val Asn Phe Val Ser Tyr Val Gln 130 135 140 ctg acg tcg cgg gcg ctg ccc agc ctg acg gac agc aag ggc tcc ctg 480 Leu Thr Ser Arg Ala Leu Pro Ser Leu Thr Asp Ser Lys Gly Ser Leu 145 150 155 160 gtg gtg gtg tcc tcg ctg ctc ggc cgc gtg ccc acg tcg ttc tcc act 528 Val Val Val Ser Ser Leu Leu Gly Arg Val Pro Thr Ser Phe Ser Thr 165 170 175 ccc tac tcg gcg gcc aag ttt gcg ctg gac ggc ttc ttc ggc tcc ctg 576 Pro Tyr Ser Ala Ala Lys Phe Ala Leu Asp Gly Phe Phe Gly Ser Leu 180 185 190 cgg cgg gag ctg gac gtg cag gac gtg aac gtg gcc atc acc atg tgc 624 Arg Arg Glu Leu Asp Val Gln Asp Val Asn Val Ala Ile Thr Met Cys 195 200 205 gtc ctg ggc ctc cga gat cgc gcc tcc gcc gcc gag gca gtc agg gga 672 Val Leu Gly Leu Arg Asp Arg Ala Ser Ala Ala Glu Ala Val Arg Gly 210 215 220 gtc acg agg gtc aag gcg gcc ccg ggg ccc aag gca gcc ctg gcc gtg 720 Val Thr Arg Val Lys Ala Ala Pro Gly Pro Lys Ala Ala Leu Ala Val 225 230 235 240 atc cgc ggc ggc gcc acg cgc gcg gcc ggc gtc ttc tac ccg tgg cgt 768 Ile Arg Gly Gly Ala Thr Arg Ala Ala Gly Val Phe Tyr Pro Trp Arg 245 250 255 ttc cgc ctg ctg tgc ttg ctc cgg cgc tgg cta ccg cgc ccg cgg gcc 816 Phe Arg Leu Leu Cys Leu Leu Arg Arg Trp Leu Pro Arg Pro Arg Ala 260 265 270 tgg ttt atc cgc cag gag ctc aac gtc acg gcc gcg gca gcc tga 861 Trp Phe Ile Arg Gln Glu Leu Asn Val Thr Ala Ala Ala Ala * 275 280 285 125 206 PRT Artificial Sequence Amino acid consensus sequence 125 Lys Val Ala Leu Val Thr Gly Ala Ser Ser Gly Ile Gly Leu Ala Ile 1 5 10 15 Ala Lys Arg Leu Ala Lys Glu Gly Ala Lys Val Val Val Ala Asp Arg 20 25 30 Asn Glu Glu Lys Leu Glu Lys Gly Ala Val Ala Lys Glu Leu Lys Glu 35 40 45 Leu Gly Gly Asn Asp Lys Asp Arg Ala Leu Ala Ile Gln Leu Asp Val 50 55 60 Thr Asp Glu Glu Ser Val Lys Ala Ala Val Glu Gln Ala Val Glu Arg 65 70 75 80 Leu Gly Arg Gly Leu Asp Val Leu Val Asn Asn Ala Gly Gly Ile Ile 85 90 95 Leu Leu Arg Pro Gly Pro Phe Ala Glu Leu Ser Arg Thr Met Glu Glu 100 105 110 Asp Trp Asp Arg Val Ile Asp Val Asn Leu Thr Gly Val Phe Leu Leu 115 120 125 Thr Arg Ala Val Leu Pro Leu Met Ala Met Lys Lys Arg Gly Gly Gly 130 135 140 Arg Ile Val Asn Ile Ser Ser Val Ala Gly Arg Lys Glu Gly Gly Leu 145 150 155 160 Val Gly Val Pro Gly Gly Ser Ala Tyr Ser Ala Ser Lys Ala Ala Val 165 170 175 Ile Gly Leu Thr Arg Ser Leu Ala Leu Glu Leu Ala Pro His Gly Gly 180 185 190 Ile Arg Val Asn Ala Val Ala Pro Gly Gly Val Asp Thr Asp 195 200 205 126 7 PRT Artificial Sequence Co-enzyme binding pattern 126 Gly Xaa Xaa Xaa Gly Xaa Gly 1 5 127 4 PRT Artificial Sequence SDR active-site pattern 127 Tyr Xaa Xaa Lys 1 128 11 PRT Artificial Sequence Short chain dehydrogenase family signature 128 Tyr Ser Ala Ala Lys Phe Ala Leu Asp Gly Phe 1 5 10 129 839 DNA Homo sapiens CDS (44)...(649) 129 cgcgagcgcg ggggccgacg ggtcgccgct gcgccgggcc ggg atg gcg gcc acc 55 Met Ala Ala Thr 1 gcg ctg ctg gag gcc ggc ctg gcg cgg gtg ctc ttc tac ccg acg ctg 103 Ala Leu Leu Glu Ala Gly Leu Ala Arg Val Leu Phe Tyr Pro Thr Leu 5 10 15 20 ctc tac acc ctg ttc cgc ggg aag gtg ccg ggt cgg gcg cac cgg gac 151 Leu Tyr Thr Leu Phe Arg Gly Lys Val Pro Gly Arg Ala His Arg Asp 25 30 35 tgg tac cac cgc atc gac ccc acc gtg ctg ctg ggc gcg ctg ccg ttg 199 Trp Tyr His Arg Ile Asp Pro Thr Val Leu Leu Gly Ala Leu Pro Leu 40 45 50 cgg agc ttg acg cgc cag ctg gta cag gac gag aac gtg cgc ggg gtg 247 Arg Ser Leu Thr Arg Gln Leu Val Gln Asp Glu Asn Val Arg Gly Val 55 60 65 atc acc atg aac gag gag tac gag acg agg ttc ctg tgc aac tct tca 295 Ile Thr Met Asn Glu Glu Tyr Glu Thr Arg Phe Leu Cys Asn Ser Ser 70 75 80 cag gag tgg aag aga cta gga gtc gag cag ctg cgg ctc agc aca gta 343 Gln Glu Trp Lys Arg Leu Gly Val Glu Gln Leu Arg Leu Ser Thr Val 85 90 95 100 gac atg act ggg atc ccc acc ttg gac aac ctc cag aag gga gtc caa 391 Asp Met Thr Gly Ile Pro Thr Leu Asp Asn Leu Gln Lys Gly Val Gln 105 110 115 ttt gct ctc aag tac cag tcg ctg ggc cag tgt gtt tac gtg cat tgt 439 Phe Ala Leu Lys Tyr Gln Ser Leu Gly Gln Cys Val Tyr Val His Cys 120 125 130 aag gct ggg cgc tcc agg agt gcc act atg gtg gca gca tac ctg att 487 Lys Ala Gly Arg Ser Arg Ser Ala Thr Met Val Ala Ala Tyr Leu Ile 135 140 145 cag gtg cac aaa tgg agt cca gag gag gct gta aga gcc atc gcc aag 535 Gln Val His Lys Trp Ser Pro Glu Glu Ala Val Arg Ala Ile Ala Lys 150 155 160 atc cgg tca tac atc cac atc agg cct ggc cag ctg gat gtt ctt aaa 583 Ile Arg Ser Tyr Ile His Ile Arg Pro Gly Gln Leu Asp Val Leu Lys 165 170 175 180 gag ttc cac aag cag att act gca cgg gca aca aag gat ggg act ttt 631 Glu Phe His Lys Gln Ile Thr Ala Arg Ala Thr Lys Asp Gly Thr Phe 185 190 195 gtc att tca aag aca tga tgtatgggga ttagaaagaa ctcaagacac 679 Val Ile Ser Lys Thr * 200 tcctgcttga tacagaacaa aaagagctta acaggaccaa cagggcttaa gcccagactt 739 gacgtaacag aaatgtgcca ataggtaata ggtaattttt ctttctctga cttgttttgt 799 tttcttgaaa taacactgtt gtgtggctag aaaaaaaaaa 839 130 201 PRT Homo sapiens 130 Met Ala Ala Thr Ala Leu Leu Glu Ala Gly Leu Ala Arg Val Leu Phe 1 5 10 15 Tyr Pro Thr Leu Leu Tyr Thr Leu Phe Arg Gly Lys Val Pro Gly Arg 20 25 30 Ala His Arg Asp Trp Tyr His Arg Ile Asp Pro Thr Val Leu Leu Gly 35 40 45 Ala Leu Pro Leu Arg Ser Leu Thr Arg Gln Leu Val Gln Asp Glu Asn 50 55 60 Val Arg Gly Val Ile Thr Met Asn Glu Glu Tyr Glu Thr Arg Phe Leu 65 70 75 80 Cys Asn Ser Ser Gln Glu Trp Lys Arg Leu Gly Val Glu Gln Leu Arg 85 90 95 Leu Ser Thr Val Asp Met Thr Gly Ile Pro Thr Leu Asp Asn Leu Gln 100 105 110 Lys Gly Val Gln Phe Ala Leu Lys Tyr Gln Ser Leu Gly Gln Cys Val 115 120 125 Tyr Val His Cys Lys Ala Gly Arg Ser Arg Ser Ala Thr Met Val Ala 130 135 140 Ala Tyr Leu Ile Gln Val His Lys Trp Ser Pro Glu Glu Ala Val Arg 145 150 155 160 Ala Ile Ala Lys Ile Arg Ser Tyr Ile His Ile Arg Pro Gly Gln Leu 165 170 175 Asp Val Leu Lys Glu Phe His Lys Gln Ile Thr Ala Arg Ala Thr Lys 180 185 190 Asp Gly Thr Phe Val Ile Ser Lys Thr 195 200 131 606 DNA Homo sapiens CDS (1)...(606) 131 atg gcg gcc acc gcg ctg ctg gag gcc ggc ctg gcg cgg gtg ctc ttc 48 Met Ala Ala Thr Ala Leu Leu Glu Ala Gly Leu Ala Arg Val Leu Phe 1 5 10 15 tac ccg acg ctg ctc tac acc ctg ttc cgc ggg aag gtg ccg ggt cgg 96 Tyr Pro Thr Leu Leu Tyr Thr Leu Phe Arg Gly Lys Val Pro Gly Arg 20 25 30 gcg cac cgg gac tgg tac cac cgc atc gac ccc acc gtg ctg ctg ggc 144 Ala His Arg Asp Trp Tyr His Arg Ile Asp Pro Thr Val Leu Leu Gly 35 40 45 gcg ctg ccg ttg cgg agc ttg acg cgc cag ctg gta cag gac gag aac 192 Ala Leu Pro Leu Arg Ser Leu Thr Arg Gln Leu Val Gln Asp Glu Asn 50 55 60 gtg cgc ggg gtg atc acc atg aac gag gag tac gag acg agg ttc ctg 240 Val Arg Gly Val Ile Thr Met Asn Glu Glu Tyr Glu Thr Arg Phe Leu 65 70 75 80 tgc aac tct tca cag gag tgg aag aga cta gga gtc gag cag ctg cgg 288 Cys Asn Ser Ser Gln Glu Trp Lys Arg Leu Gly Val Glu Gln Leu Arg 85 90 95 ctc agc aca gta gac atg act ggg atc ccc acc ttg gac aac ctc cag 336 Leu Ser Thr Val Asp Met Thr Gly Ile Pro Thr Leu Asp Asn Leu Gln 100 105 110 aag gga gtc caa ttt gct ctc aag tac cag tcg ctg ggc cag tgt gtt 384 Lys Gly Val Gln Phe Ala Leu Lys Tyr Gln Ser Leu Gly Gln Cys Val 115 120 125 tac gtg cat tgt aag gct ggg cgc tcc agg agt gcc act atg gtg gca 432 Tyr Val His Cys Lys Ala Gly Arg Ser Arg Ser Ala Thr Met Val Ala 130 135 140 gca tac ctg att cag gtg cac aaa tgg agt cca gag gag gct gta aga 480 Ala Tyr Leu Ile Gln Val His Lys Trp Ser Pro Glu Glu Ala Val Arg 145 150 155 160 gcc atc gcc aag atc cgg tca tac atc cac atc agg cct ggc cag ctg 528 Ala Ile Ala Lys Ile Arg Ser Tyr Ile His Ile Arg Pro Gly Gln Leu 165 170 175 gat gtt ctt aaa gag ttc cac aag cag att act gca cgg gca aca aag 576 Asp Val Leu Lys Glu Phe His Lys Gln Ile Thr Ala Arg Ala Thr Lys 180 185 190 gat ggg act ttt gtc att tca aag aca tga 606 Asp Gly Thr Phe Val Ile Ser Lys Thr * 195 200 132 173 PRT Artificial Sequence Amino acid consensus sequence 132 Gly Pro Ser Glu Ile Leu Pro His Leu Tyr Leu Gly Ser Tyr Ser Thr 1 5 10 15 Ala Ser Glu Ala Asn Leu Ala Leu Leu Lys Lys Leu Gly Ile Thr His 20 25 30 Val Ile Asn Val Thr Glu Glu Val Pro Asn Pro Phe Glu Leu Asp Lys 35 40 45 Lys Asn Asp Arg His Tyr Thr Asn Ala Tyr Ile Ser Lys Asn Ser Gly 50 55 60 Phe Thr Tyr Leu Gln Ile Pro Asn Val Asp Asp His Ile Tyr Tyr His 65 70 75 80 Ile Ala Trp Asn His Glu Thr Lys Ile Ser Lys Tyr Phe Asp Glu Ala 85 90 95 Val Asp Phe Ile Asp Asp Ala Arg Gln Lys Gly Gly Lys Val Leu Val 100 105 110 His Cys Gln Ala Gly Ile Ser Arg Ser Ala Thr Leu Ile Ile Ala Tyr 115 120 125 Leu Met Lys Thr Arg Asn Leu Ser Leu Asn Glu Ala Tyr Asp Phe Val 130 135 140 Tyr Val Tyr His Ile Lys Glu Arg Arg Cys Pro Ile Ile Ser Pro Asn 145 150 155 160 Phe Gly Phe Leu Arg Gln Leu Ile Glu Tyr Glu Arg Lys 165 170 133 172 PRT Artificial Sequence Amino acid consensus sequence 133 Gly Pro Ser Glu Ile Leu Pro His Leu Tyr Leu Gly Ser Tyr Ser Asp 1 5 10 15 Ala Ser Glu Ala Asn Leu Ala Leu Leu Lys Lys Leu Gly Ile Thr His 20 25 30 Val Ile Asn Val Thr Glu Glu Val Pro Asn Asn Phe Glu Leu Lys Lys 35 40 45 Lys Asn Asp Arg Tyr Tyr Thr Asn Glu Tyr Ile Ser Lys Gly Ser Gly 50 55 60 Phe Thr Tyr Leu Gln Ile Pro Asn Val Asp Asp Ile Tyr Tyr His Ile 65 70 75 80 Ala Trp Asn Thr Glu Thr Lys Ile Ser Lys Tyr Leu Glu Glu Ala Val 85 90 95 Glu Phe Ile Glu Asp Ala Glu Lys Lys Gly Gly Lys Val Leu Val His 100 105 110 Cys Gln Ala Gly Val Ser Arg Ser Ala Thr Leu Val Ile Ala Tyr Leu 115 120 125 Met Lys Thr Arg Asn Leu Ser Leu Arg Asp Ala Tyr Asp Phe Val Tyr 130 135 140 Val Tyr His Ile Lys Glu Arg Arg Cys Pro Ile Ile Ser Pro Asn Phe 145 150 155 160 Gly Phe Leu Arg Gln Leu Ile Glu Tyr Glu Arg Lys 165 170 134 13 PRT Artificial Sequence Tyrosine specific protein phosphatase active site signature 134 Xaa His Cys Xaa Xaa Gly Xaa Xaa Xaa Xaa Xaa Xaa Xaa 1 5 10 135 7 PRT Artificial Sequence Active site motif of the tyrosine phosphatase signature 135 Cys Xaa Xaa Xaa Xaa Xaa Arg 1 5 136 21 PRT Artificial Sequence Dual specificity phosphatase extended active site signature 136 Val Xaa Val His Cys Xaa Xaa Gly Xaa Ser Arg Ser Xaa Thr Xaa Xaa 1 5 10 15 Xaa Ala Tyr Xaa Met 20

Claims

What is claimed is:

1. An isolated 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 nucleic acid molecule selected from the group consisting of:

a) a nucleic acid molecule comprising a nucleotide sequence which is at least 60% identical to the nucleotide sequence of SEQ ID NO:1, 3, 4, 6, 7, 9, 10, 12, 13, 15, 16, 18, 53, 55, 61, 63, 67, 69, 78, 80, 88, 90, 100, 102, 113, 115, 122, 124, 129 or 131, or the nucleotide sequence of the DNA insert of the plasmid deposited with ATCC Accession Number PTA-3437 or PTA-3435;

b) a nucleic acid molecule comprising a fragment of at least 15 nucleotides of the nucleotide sequence of SEQ ID NO:1, 3, 4, 6, 7, 9, 10, 12, 13, 15, 16, 18, 53, 55, 61, 63, 67, 69, 78, 80, 88, 90, 100, 102, 113, 115, 122, 124, 129 or 131, or the nucleotide sequence of the DNA insert of the plasmid deposited with ATCC Accession Number PTA-3437 or PTA-3435;

c) a nucleic acid molecule which encodes a polypeptide comprising the amino acid sequence of SEQ ID NO:2, 5, 8, 11, 14, 17, 54, 62, 68, 79, 89, 101, 114, 123 or 130, or the amino acid sequence encoded by the cDNA insert of the plasmid deposited with ATCC Accession Number PTA-3437 or PTA-3435;

d) a nucleic acid molecule which encodes a fragment of a polypeptide comprising the amino acid sequence of SEQ ID NO:2, 5, 8, 11, 14, 17, 54, 62, 68, 79, 89, 101, 114, 123 or 130, or the amino acid sequence encoded by the cDNA insert of the plasmid deposited with ATCC Accession Number PTA-3437 or PTA-3435, wherein the fragment comprises at least 15 contiguous amino acids of SEQ ID NO:2, 5, 8, 11, 14, 17, 54, 62, 68, 79, 89, 101, 114, 123 or 130, or the amino acid sequence encoded by the cDNA insert of the plasmid deposited with ATCC Accession Number PTA-3437 or PTA-3435;

e) a nucleic acid molecule which encodes a naturally occurring allelic variant of a polypeptide comprising the amino acid sequence of SEQ ID NO:2, 5, 8, 11, 14, 17, 54, 62, 68, 79, 89, 101, 114, 123 or 130, or the amino acid sequence encoded by the cDNA insert of the plasmid deposited with ATCC Accession Number PTA-3437 or PTA-3435, wherein the nucleic acid molecule hybridizes to a nucleic acid molecule comprising SEQ ID NO:1, 3, 4, 6, 7, 9, 10, 12, 13, 15, 16, 18, 53, 55, 61, 63, 67, 69, 78, 80, 88, 90, 100, 102, 113, 115, 122, 124, 129 or 131, or a complement thereof, under stringent conditions;

f) a nucleic acid molecule comprising the nucleotide sequence of SEQ ID NO:1, 3, 4, 6, 7, 9, 10, 12, 13, 15, 16, 18, 53, 55, 61, 63, 67, 69, 78, 80, 88, 90, 100, 102, 113, 115, 122, 124, 129 or 131, or the nucleotide sequence of the DNA insert of the plasmid deposited with ATCC Accession Number PTA-3437 or PTA-3435; and

g) a nucleic acid molecule which encodes a polypeptide comprising the amino acid sequence of SEQ ID NO:2, 5, 8, 11, 14, 17, 54, 62, 68, 79, 89, 101, 114, 123 or 130, or the amino acid sequence encoded by the cDNA insert of the plasmid deposited with ATCC Accession Number PTA-3437 or PTA-3435.

2. The isolated nucleic acid molecule of claim 1, which is the nucleotide sequence SEQ ID NO:1, 4, 7, 10, 13, 16, 53, 61, 67, 78, 88, 100, 113, 122or 129.

3. A host cell which contains the nucleic acid molecule of claim 1.

4. An isolated 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 polypeptide selected from the group consisting of:

a) a polypeptide which is encoded by a nucleic acid molecule comprising a nucleotide sequence which is at least 60% identical to a nucleic acid comprising the nucleotide sequence of SEQ ID NO:1, 3, 4, 6, 7, 9, 10, 12, 13, 15, 16, 18, 53, 55, 61, 63, 67, 69, 78, 80, 88, 90, 100, 102, 113, 115, 122, 124, 129 or 131, or the nucleotide sequence of the DNA insert of the plasmid deposited with ATCC Accession Number PTA-3437 or PTA-3435, or a complement thereof;

b) a naturally occurring allelic variant of a polypeptide comprising the amino acid sequence of SEQ ID NO:2, 5, 8, 11, 14, 17, 54, 62, 68, 79, 89, 101, 114, 123 or 130, or the amino acid sequence encoded by the cDNA insert of the plasmid deposited with ATCC Accession Number PTA-3437 or PTA-3435, wherein the polypeptide is encoded by a nucleic acid molecule which hybridizes to a nucleic acid molecule comprising SEQ ID NO:1, 3, 4, 6, 7, 9, 10, 12, 13, 15, 16, 18, 53, 55, 61, 63, 67, 69, 78, 80, 88, 90, 100, 102, 113, 115, 122, 124, 129 or 131, or a complement thereof under stringent conditions;

c) a fragment of a polypeptide comprising the amino acid sequence of SEQ ID NO:2, 5, 8, 11, 14, 17, 54, 62, 68, 79, 89, 101, 114, 123 or 130, or the amino acid sequence encoded by the cDNA insert of the plasmid deposited with ATCC Accession Number PTA-3437 or PTA-3435, wherein the fragment comprises at least 15 contiguous amino acids of SEQ ID NO:2, 5, 8, 11, 14, 17, 54, 62, 68, 79, 89, 101, 114, 123 or 130; and

d) the amino acid sequence of SEQ ID NO:2, 5, 8, 11, 14, 17, 54, 62, 68, 79, 89, 101, 114, 123 or 130.

5. An antibody which selectively binds to a polypeptide of claim 4.

6. The polypeptide of claim 4, further comprising heterologous amino acid sequences.

7. A method for producing a polypeptide selected from the group consisting of:

a) a polypeptide comprising the amino acid sequence of SEQ ID NO:2, 5, 8, 11, 14, 17, 54, 62, 68, 79, 89, 101, 114, 123 or 130, or the amino acid sequence encoded by the cDNA insert of the plasmid deposited with ATCC Accession Number PTA-3437 or PTA-3435;

b) a polypeptide comprising a fragment of the amino acid sequence of SEQ ID NO:2, 5, 8, 11, 14, 17, 54, 62, 68, 79, 89, 101, 114, 123 or 130, or the amino acid sequence encoded by the cDNA insert of the plasmid deposited with ATCC Accession Number PTA-3437 or PTA-3435, wherein the fragment comprises at least 15 contiguous amino acids of SEQ ID NO:2, 5, 8, 11, 14, 17, 54, 62, 68, 79, 89, 101, 114, 123 or 130, or the amino acid sequence encoded by the cDNA insert of the plasmid deposited with ATCC Accession Number PTA-3437 or PTA-3435;

c) a naturally occurring allelic variant of a polypeptide comprising the amino acid sequence of SEQ ID NO:2, 5, 8, 11, 14, 17, 54, 62, 68, 79, 89, 101, 114, 123 or 130, or the amino acid sequence encoded by the cDNA insert of the plasmid deposited with ATCC Accession Number PTA-3437 or PTA-3435, wherein the polypeptide is encoded by a nucleic acid molecule which hybridizes to a nucleic acid molecule comprising SEQ ID NO:1, 3, 4, 6, 7, 9, 10, 12, 13, 15, 16, 18, 53, 55, 61, 63, 67, 69, 78, 80, 88, 90, 100, 102, 113, 115, 122, 124, 129 or 131; and

d) the amino acid sequence of SEQ ID NO:2, 5, 8, 11, 14, 17, 54, 62, 68, 79, 89, 101, 114, 123 or 130;

comprising culturing the host cell of claim 3 under conditions in which the nucleic acid molecule is expressed.

8. A method for detecting the presence of a nucleic acid molecule of claim 1 or a polypeptide encoded by the nucleic acid molecule in a sample, comprising:

a) contacting the sample with a compound which selectively hybridizes to the nucleic acid molecule of claim 1 or binds to the polypeptide encoded by the nucleic acid molecule; and

b) determining whether the compound hybridizes to the nucleic acid or binds to the polypeptide in the sample.

9. A kit comprising a compound which selectively hybridizes to a nucleic acid molecule of claim 1 or binds to a polypeptide encoded by the nucleic acid molecule and instructions for use.

10. A method for identifying a compound which binds to a polypeptide or modulates the activity of the polypeptide of claim 4 comprising the steps of:

a) contacting a polypeptide, or a cell expressing a polypeptide of claim 4 with a test compound; and

b) determining whether the polypeptide binds to the test compound or determining the effect of the test compound on the activity of the polypeptide.

11. A method for modulating the activity of a polypeptide of claim 4 comprising contacting the polypeptide or a cell expressing the polypeptide with a compound which binds to the polypeptide in a sufficient concentration to modulate the activity of the polypeptide.

12. A method for identifying a compound capable of treating a disorder characterized by aberrant 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 activity, comprising assaying the ability of the compound to modulate 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 nucleic acid expression or 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 polypeptide activity, thereby identifying a compound capable of treating a disorder characterized by aberrant 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 activity.

13. A method of identifying a nucleic acid molecule associated with a disorder characterized by aberrant 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 activity, comprising:

a) contacting a sample from a subject with a disorder characterized by aberrant 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 activity, comprising nucleic acid molecules with a hybridization probe comprising at least 25 contiguous nucleotides of SEQ ID NO:1, 4, 7, 10, 13, 16, 53, 61, 67, 78, 88, 100, 113, 122 or 129 defined in claim 2; and

b) detecting the presence of a nucleic acid molecule in the sample that hybridizes to the probe, thereby identifying a nucleic acid molecule associated with a disorder characterized by aberrant 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 activity.

14. A method of identifying a polypeptide associated with a disorder characterized by aberrant 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 activity, comprising:

a) contacting a sample comprising polypeptides with a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 polypeptide defined in claim 4; and

b) detecting the presence of a polypeptide in the sample that binds to the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 binding partner, thereby identifying the polypeptide associated with a disorder characterized by aberrant 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 activity.

15. A method of identifying a subject having a disorder characterized by aberrant 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 activity, comprising:

a) contacting a sample obtained from the subject comprising nucleic acid molecules with a hybridization probe comprising at least 25 contiguous nucleotides of SEQ ID NO:1, 4, 7, 10, 13, 16, 53, 61, 67, 78, 88, 100, 113, 122 or 129 defined in claim 2; and

b) detecting the presence of a nucleic acid molecule in the sample that hybridizes to the probe, thereby identifying a subject having a disorder characterized by aberrant 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 activity.

16. A method for treating a subject having a disorder characterized by aberrant 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 activity, or a subject at risk of developing a disorder characterized by aberrant 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 activity, comprising administering to the subject a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 modulator of the nucleic acid molecule defined in claim 1 or the polypeptide encoded by the nucleic acid molecule or contacting a cell with a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 modulator.

17. The method defined in claim 16 wherein said disorder is a cellular proliferative and/or differentiative disorder, brain disorder, platelet disorder, breast disorder, colon disorder, kidney (renal) disorder, lung disorder, ovarian disorder, prostate disorder, hematopoeitic disorder, pancreatic disorder, skeletal muscle disorder, skin (dermal) disorder, disorder associated with bone metabolism, immune, e.g., inflammatory, disorder, cardiovascular disorder, endothelial cell disorder, liver disorder, viral diseases, pain disorder, metabolic disorder, neurological or CNS disorder, erythroid disorder or anemic disorder.

18. The method of claim 16, wherein the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 modulator is

a) a small molecule;

b) peptide;

c) phosphopeptide;

d) anti-26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 antibody;

e) a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 polypeptide comprising the amino acid sequence of SEQ ID NO:2, 5, 8, 11, 14, 17, 54, 62, 68, 79, 89, 101, 114, 123 or 130, or a fragment thereof;

f) a 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 polypeptide comprising an amino acid sequence which is at least 90 percent identical to the amino acid sequence of SEQ ID NO:2, 5, 8, 11, 14, 17, 54, 62, 68, 79, 89, 101, 114, 123 or 130, wherein the percent identity is calculated using the ALIGN program for comparing amino acid sequences, a PAM120 weight residue table, a gap length penalty of 12, and a gap penalty of 4; or

g) an isolated naturally occurring allelic variant of a polypeptide consisting of the amino acid sequence of SEQ ID NO:2, 5, 8, 11, 14, 17, 54, 62, 68, 79, 89, 101, 114, 123 or 130, wherein the polypeptide is encoded by a nucleic acid molecule which hybridizes to a complement of a nucleic acid molecule consisting of SEQ ID NO:1, 4, 7, 10, 13, 16, 53, 61, 67, 78, 88, 100, 113, 122 or 129 at 6×SSC at 45° C., followed by one or more washes in 0.2×SSC, 0.1% SDS at 65° C.

19. The method of claim 16, wherein the 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 modulator is

a) an antisense 26199, 33530, 33949, 47148, 50226, 58764, 62113, 32144, 32235, 23565, 13305, 14911, 86216, 25206 or 8843 nucleic acid molecule;

b) is a ribozyme;

c) the nucleotide sequence of SEQ ID NO:1, 4, 7, 10, 13, 16, 53, 61, 67, 78, 88, 100, 113, 122 or 129 or a fragment thereof;

d) a nucleic acid molecule encoding a polypeptide comprising an amino acid sequence which is at least 90 percent identical to the amino acid sequence of SEQ ID NO:2, 5, 8, 11, 14, 17, 54, 62, 68, 79, 89, 101, 114, 123 or 130, wherein the percent identity is calculated using the ALIGN program for comparing amino acid sequences, a PAM120 weight residue table, a gap length penalty of 12, and a gap penalty of 4;

e) a nucleic acid molecule encoding a naturally occurring allelic variant of a polypeptide comprising the amino acid sequence of SEQ ID NO:2, 5, 8, 11, 14, 17, 54, 62, 68, 79, 89, 101, 114, 123 or 130, wherein the nucleic acid molecule which hybridizes to a complement of a nucleic acid molecule consisting of SEQ ID NO:1, 4, 7, 10, 13, 16, 53, 61, 67, 78, 88, 100, 113, 122 or 129 at 6×SSC at 45° C., followed by one or more washes in 0.2×SSC, 0.1% SDS at 65° C.; or

f) a gene therapy vector.