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WO2001060840A3 - Complementation assay assessing protein solubility and folding - Google Patents

Complementation assay assessing protein solubility and folding Download PDF

Info

Publication number
WO2001060840A3
WO2001060840A3 PCT/US2001/005097 US0105097W WO0160840A3 WO 2001060840 A3 WO2001060840 A3 WO 2001060840A3 US 0105097 W US0105097 W US 0105097W WO 0160840 A3 WO0160840 A3 WO 0160840A3
Authority
WO
WIPO (PCT)
Prior art keywords
protein
folding
improper
marker protein
protein solubility
Prior art date
Application number
PCT/US2001/005097
Other languages
French (fr)
Other versions
WO2001060840A2 (en
Inventor
Philip Jordan Thomas
John F Hunt
William Christian Wigley
Rhesa D Stidham
Original Assignee
Univ Texas
Univ Columbia
Philip Jordan Thomas
John F Hunt
Rhesa D Stidham
William Christian Wigley
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Univ Texas, Univ Columbia, Philip Jordan Thomas, John F Hunt, Rhesa D Stidham, William Christian Wigley filed Critical Univ Texas
Priority to JP2001560224A priority Critical patent/JP4772247B2/en
Priority to EP01909281A priority patent/EP1257829B1/en
Priority to AU2001237055A priority patent/AU2001237055B2/en
Priority to CA2398834A priority patent/CA2398834C/en
Priority to DK01909281T priority patent/DK1257829T3/en
Priority to DE60123755T priority patent/DE60123755T2/en
Priority to AU3705501A priority patent/AU3705501A/en
Publication of WO2001060840A2 publication Critical patent/WO2001060840A2/en
Publication of WO2001060840A3 publication Critical patent/WO2001060840A3/en

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
    • C07K1/107—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length by chemical modification of precursor peptides
    • C07K1/113—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length by chemical modification of precursor peptides without change of the primary structure

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Biochemistry (AREA)
  • Biophysics (AREA)
  • General Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Medicinal Chemistry (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Peptides Or Proteins (AREA)
  • Investigating Or Analysing Biological Materials (AREA)

Abstract

Many proteins, when produced recombinantly, suffer from improper processing, folding and lack normal solubility. Modified proteins, including those indicative of disease states, also can have such defects. The present invention is directed to methods of identifying proper and improper protein folding, aberrant processing and/or insolubility. The method relies on the use of two components: a specialized fusion protein and structural complementation. The fusion protein contains sequences from the protein of interest and one portion of a marker protein that, by itself, is not active. A host cell then provides the remainder of the marker protein that serves to 'complement' the function of the fused marker protein such that their association restores activity, permitting detection.
PCT/US2001/005097 2000-02-14 2001-02-14 Complementation assay assessing protein solubility and folding WO2001060840A2 (en)

Priority Applications (7)

Application Number Priority Date Filing Date Title
JP2001560224A JP4772247B2 (en) 2000-02-14 2001-02-14 Complementary assays to assess protein solubility and folding
EP01909281A EP1257829B1 (en) 2000-02-14 2001-02-14 Complementation assay assessing protein solubility and folding
AU2001237055A AU2001237055B2 (en) 2000-02-14 2001-02-14 Complementation assay assessing protein solubility and folding
CA2398834A CA2398834C (en) 2000-02-14 2001-02-14 Protein/solubility folding assessed by structural complementation
DK01909281T DK1257829T3 (en) 2000-02-14 2001-02-14 Complementation assay assessing protein solubility and folding
DE60123755T DE60123755T2 (en) 2000-02-14 2001-02-14 COMPLEMENTATION ASSAY FOR DETECTING PROTEIN SOLUBILITY AND FOLDING
AU3705501A AU3705501A (en) 2000-02-14 2001-02-14 Protein/solubility folding assessed by structural complementation

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
US18228300P 2000-02-14 2000-02-14
US60/182,283 2000-02-14
US77505100A 2001-01-31 2001-01-31
US09/775,051 2001-01-31

Publications (2)

Publication Number Publication Date
WO2001060840A2 WO2001060840A2 (en) 2001-08-23
WO2001060840A3 true WO2001060840A3 (en) 2002-04-25

Family

ID=22667801

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2001/005097 WO2001060840A2 (en) 2000-02-14 2001-02-14 Complementation assay assessing protein solubility and folding

Country Status (1)

Country Link
WO (1) WO2001060840A2 (en)

Families Citing this family (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR2883886B1 (en) * 2005-04-05 2010-12-03 Biomethodes METHOD OF OBTAINING SOLUBLE OR BETTER EXPRESS VARIANTS OF A PROTEIN OF INTEREST
US8541175B2 (en) 2006-03-13 2013-09-24 The Board Of Trustees Of The Leland Stanford Junior University Detection of molecular interactions using a reduced affinity enzyme complementation reporter system
US8101373B2 (en) 2007-10-12 2012-01-24 Discoverx Corporation β-galactosidase donor fragments
WO2011017319A1 (en) 2009-08-03 2011-02-10 University Of Pittsburgh - Of The Commonwealth System Of Higher Education Methods of treating disorders associated with protein polymerization
US8809617B2 (en) * 2009-11-05 2014-08-19 The University of Pittsburgh—Of the Commonwealth System of Higher Education Automated high-content live animal drug screening using C. elegans
US9072772B2 (en) 2009-11-05 2015-07-07 University of Pittsburgh—of the Commonwealth System of Higher Education Methods of treating disorders associated with protein aggregation
US8569057B2 (en) 2011-06-23 2013-10-29 Discoverx Corporation Monitoring protein trafficking using beta-galactosidase reporter fragment complementation

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1998034120A1 (en) * 1997-01-31 1998-08-06 Universite De Montreal Protein fragment complementation assays to detect biomolecular interactions
WO1998044350A1 (en) * 1997-04-02 1998-10-08 The Board Of Trustees Of The Leland Stanford Junior University Detection of molecular interactions by reporter subunit complementation

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1998034120A1 (en) * 1997-01-31 1998-08-06 Universite De Montreal Protein fragment complementation assays to detect biomolecular interactions
WO1998044350A1 (en) * 1997-04-02 1998-10-08 The Board Of Trustees Of The Leland Stanford Junior University Detection of molecular interactions by reporter subunit complementation

Non-Patent Citations (6)

* Cited by examiner, † Cited by third party
Title
ABBAS-TERKI TOUFIK ET AL: "alpha-complemented beta-galactosidase. An in vivo model substrate for the molecular chaperone heat-shock protein 90 in yeast.", EUROPEAN JOURNAL OF BIOCHEMISTRY, vol. 266, no. 2, December 1999 (1999-12-01), pages 517 - 523, XP002182770, ISSN: 0014-2956 *
CAMPBELL VALOIS F -X ET AL: "Utilisation of the PCA strategy to study the folding of the RBD of raf.", FASEB JOURNAL, vol. 13, no. 7, 23 April 1999 (1999-04-23), Annual Meeting of the American Societies for Experimental Biology on Biochemistry and Molecular Biology 99;San Francisco, California, USA; May 16-20, 1999, pages A1387, XP002182768, ISSN: 0892-6638 *
JOHNSSON N ET AL: "SPLIT UBIQUITIN AS A SENSOR OF PROTEIN INTERACTIONS INV VIVO", PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF USA, NATIONAL ACADEMY OF SCIENCE. WASHINGTON, US, vol. 91, 1 October 1994 (1994-10-01), pages 10340 - 10344, XP002064564, ISSN: 0027-8424 *
KO Y H ET AL: "THE CYSTIC FIBROSIS TRANSMEMBRANE CONDUCTANCE REGULATOR. OVEREXPRESSION, PURIFICATION, AND CHARACTERIZATION OF WILD TYPE ANDDELTAF508 MUTANT FORMS OF THE FIRST NUCLEOTIDE BINDING FOLD IN FUSION WITH THE MALTOSE-BINDING PROTEIN", CELL, CELL PRESS, CAMBRIDGE, NA, US, vol. 268, no. 32, 15 November 1993 (1993-11-15), pages 24330 - 24338, XP002055212, ISSN: 0092-8674 *
NIXON ANDREW E ET AL: "Improvement in the efficiency of formyl transfer of a GAR transformylase hybrid enzyme.", PROTEIN ENGINEERING, vol. 13, no. 5, May 2000 (2000-05-01), pages 323 - 327, XP002182771, ISSN: 0269-2139 *
WALDO GEOFFREY S ET AL: "Rapid protein-folding assay using green fluorescent protein", NATURE BIOTECHNOLOGY, vol. 17, July 1999 (1999-07-01), pages 691 - 695, XP002182769 *

Also Published As

Publication number Publication date
WO2001060840A2 (en) 2001-08-23

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