Super Spy variants implicate flexibility in chaperone action

Elife. 2014:3:e01584. doi: 10.7554/eLife.01584. Epub 2014 Feb 4.

Abstract

Experimental study of the role of disorder in protein function is challenging. It has been proposed that proteins utilize disordered regions in the adaptive recognition of their various binding partners. However apart from a few exceptions, defining the importance of disorder in promiscuous binding interactions has proven to be difficult. In this paper, we have utilized a genetic selection that links protein stability to antibiotic resistance to isolate variants of the newly discovered chaperone Spy that show an up to 7 fold improved chaperone activity against a variety of substrates. These "Super Spy" variants show tighter binding to client proteins and are generally more unstable than is wild type Spy and show increases in apparent flexibility. We establish a good relationship between the degree of their instability and the improvement they show in their chaperone activity. Our results provide evidence for the importance of disorder and flexibility in chaperone function. DOI: http://dx.doi.org/10.7554/eLife.01584.001.

Keywords: binding; chaperones; intrinsic disorder; protein folding; protein stability.

Publication types

  • Comparative Study
  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anti-Bacterial Agents / pharmacology
  • Binding Sites
  • Cross-Linking Reagents / chemistry
  • Drug Resistance, Bacterial / genetics
  • Escherichia coli / drug effects
  • Escherichia coli / genetics
  • Escherichia coli / growth & development
  • Escherichia coli / metabolism*
  • Escherichia coli Proteins / chemistry
  • Escherichia coli Proteins / genetics
  • Escherichia coli Proteins / metabolism*
  • Genetic Variation
  • Genotype
  • Hydrophobic and Hydrophilic Interactions
  • Microbial Viability
  • Molecular Chaperones / chemistry
  • Molecular Chaperones / genetics
  • Molecular Chaperones / metabolism*
  • Molecular Docking Simulation
  • Mutation
  • Periplasmic Proteins / chemistry
  • Periplasmic Proteins / genetics
  • Periplasmic Proteins / metabolism*
  • Phenotype
  • Protein Binding
  • Protein Conformation
  • Protein Folding
  • Protein Stability
  • Structure-Activity Relationship

Substances

  • Anti-Bacterial Agents
  • Cross-Linking Reagents
  • Escherichia coli Proteins
  • Molecular Chaperones
  • Periplasmic Proteins
  • Spy protein, E coli