Activation and inhibition of the receptor histidine kinase AgrC occurs through opposite helical transduction motions

Mol Cell. 2014 Mar 20;53(6):929-40. doi: 10.1016/j.molcel.2014.02.029.

Authors

Boyuan Wang¹, Aishan Zhao², Richard P Novick³, Tom W Muir⁴

Affiliations

¹ Department of Chemistry, Princeton University, Frick Chemistry Building, Washington Road, Princeton, NJ 08544, USA; Graduate Program, The Rockefeller University, 1230 York Avenue, New York, NY 10065, USA.
² Department of Chemistry, Princeton University, Frick Chemistry Building, Washington Road, Princeton, NJ 08544, USA.
³ Skirball Institute, Department of Microbiology, NYU Medical Center, 540 First Avenue, New York, NY 10016, USA.
⁴ Department of Chemistry, Princeton University, Frick Chemistry Building, Washington Road, Princeton, NJ 08544, USA. Electronic address: muir@princeton.edu.

Abstract

Staphylococcus aureus virulence is regulated when secreted autoinducing peptides (AIPs) are recognized by a membrane-bound receptor histidine kinase (RHK), AgrC. Some AIPs are agonists of virulence gene expression, while others are antagonists. It is unclear how AIP binding regulates AgrC activity. Here, we reconstitute an AgrC family member, AgrC-I, using nanometer-scale lipid bilayer discs. We show that AgrC-I requires membranes rich in anionic lipids to function. The agonist, AIP-I, binds AgrC-I noncooperatively in a 2:2 stoichiometry, while an antagonist ligand, AIP-II, functions as an inverse agonist of the kinase activity. We also demonstrate the kinase and sensor domains in AgrC are connected by a helical linker whose conformational state exercises rheostat-like control over the kinase activity. Binding of agonist or inverse-agonist peptides results in twisting of the linker in different directions. These two observations provide a view of the molecular motions triggered by ligand binding in an intact membrane-bound RHK.

Publication types

Research Support, N.I.H., Extramural

MeSH terms

Amino Acid Sequence
Bacterial Proteins / chemistry
Bacterial Proteins / genetics*
Bacterial Proteins / metabolism
Binding Sites
Escherichia coli / genetics
Escherichia coli / metabolism
Gene Expression Regulation, Bacterial*
Ligands
Lipid Bilayers / chemistry*
Models, Biological
Molecular Sequence Data
Peptides, Cyclic / chemistry
Peptides, Cyclic / genetics*
Peptides, Cyclic / metabolism
Phospholipids / chemistry
Protein Binding
Protein Isoforms / chemistry
Protein Isoforms / genetics
Protein Isoforms / metabolism
Protein Kinases / chemistry
Protein Kinases / genetics*
Protein Kinases / metabolism
Protein Structure, Secondary
Protein Structure, Tertiary
Recombinant Proteins / chemistry
Recombinant Proteins / genetics
Recombinant Proteins / metabolism
Sequence Alignment
Sequence Homology, Amino Acid
Signal Transduction*
Staphylococcus aureus / genetics*
Staphylococcus aureus / metabolism
Staphylococcus aureus / pathogenicity
Virulence

Substances

AgrD protein, Staphylococcus
Bacterial Proteins
Ligands
Lipid Bilayers
Peptides, Cyclic
Phospholipids
Protein Isoforms
Recombinant Proteins
Protein Kinases
AgrC protein, Staphylococcus

Abstract

Publication types

MeSH terms

Substances

Grants and funding