1887

Abstract

It has long been thought that chaperones are primarily attracted to their clients through the hydrophobic effect. However, in in vitro studies on the interaction between the chaperone Spy and its substrate Im7, we recently showed that long-range electrostatic interactions also play a key role. Spy functions in the periplasm of Gram-negative bacteria, which is surrounded by a permeable outer membrane. The ionic conditions in the periplasm therefore closely mimic those in the media, which allowed us to vary the ionic strength of the in vivo folding environment. Using folding biosensors that link protein folding to antibiotic resistance, we were able to monitor Spy chaperone activity in Escherichia coli in vivo as a function of media salt concentration. The chaperone activity of Spy decreased when the ionic strength of the media was increased, strongly suggesting that electrostatic forces play a vital role in the action of Spy in vivo.

Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.000676
2018-06-05
2019-10-15
Loading full text...

Full text loading...

/deliver/fulltext/micro/164/7/992.html?itemId=/content/journal/micro/10.1099/mic.0.000676&mimeType=html&fmt=ahah

References

  1. Balchin D, Hayer-Hartl M, Hartl FU. In vivo aspects of protein folding and quality control. Science 2016;353:aac4354 [CrossRef][PubMed]
    [Google Scholar]
  2. Kim YE, Hipp MS, Bracher A, Hayer-Hartl M, Hartl FU. Molecular chaperone functions in protein folding and proteostasis. Annu Rev Biochem 2013;82:323–355 [CrossRef][PubMed]
    [Google Scholar]
  3. Koldewey P, Horowitz S, Bardwell JCA. Chaperone-client interactions: non-specificity engenders multifunctionality. J Biol Chem 2017;292:12010–12017 [CrossRef][PubMed]
    [Google Scholar]
  4. Koldewey P, Stull F, Horowitz S, Martin R, Bardwell JCA. Forces driving chaperone action. Cell 2016;166:369–379 [CrossRef][PubMed]
    [Google Scholar]
  5. Coyle JE, Jaeger J, Gross M, Robinson CV, Radford SE. Structural and mechanistic consequences of polypeptide binding by GroEL. Fold Des 1997;2:R93–R104 [CrossRef][PubMed]
    [Google Scholar]
  6. Schreiber G, Haran G, Zhou HX. Fundamental aspects of protein-protein association kinetics. Chem Rev 2009;109:839–860 [CrossRef][PubMed]
    [Google Scholar]
  7. Selzer T, Schreiber G. Predicting the rate enhancement of protein complex formation from the electrostatic energy of interaction. J Mol Biol 1999;287:409–419 [CrossRef][PubMed]
    [Google Scholar]
  8. Ferguson N, Capaldi AP, James R, Kleanthous C, Radford SE. Rapid folding with and without populated intermediates in the homologous four-helix proteins Im7 and Im9. J Mol Biol 1999;286:1597–1608 [CrossRef][PubMed]
    [Google Scholar]
  9. Capaldi AP, Kleanthous C, Radford SE. Im7 folding mechanism: misfolding on a path to the native state. Nat Struct Biol 2002;9:209–216 [CrossRef][PubMed]
    [Google Scholar]
  10. Stull F, Koldewey P, Humes JR, Radford SE, Bardwell JCA. Substrate protein folds while it is bound to the ATP-independent chaperone Spy. Nat Struct Mol Biol 2016;23:53–58 [CrossRef][PubMed]
    [Google Scholar]
  11. Horowitz S, Salmon L, Koldewey P, Ahlstrom LS, Martin R et al. Visualizing chaperone-assisted protein folding. Nat Struct Mol Biol 2016;23:691–697 [CrossRef][PubMed]
    [Google Scholar]
  12. Quan S, Koldewey P, Tapley T, Kirsch N, Ruane KM et al. Genetic selection designed to stabilize proteins uncovers a chaperone called Spy. Nat Struct Mol Biol 2011;18:262–269 [CrossRef][PubMed]
    [Google Scholar]
  13. Foit L, Morgan GJ, Kern MJ, Steimer LR, von Hacht AA et al. Optimizing protein stability in vivo. Mol Cell 2009;36:861–871 [CrossRef][PubMed]
    [Google Scholar]
  14. Nikaido H. Molecular basis of bacterial outer membrane permeability revisited. Microbiol Mol Biol Rev 2003;67:593–656 [CrossRef][PubMed]
    [Google Scholar]
  15. Park SH, Kukushkin Y, Gupta R, Chen T, Konagai A et al. PolyQ proteins interfere with nuclear degradation of cytosolic proteins by sequestering the Sis1p chaperone. Cell 2013;154:134–145 [CrossRef][PubMed]
    [Google Scholar]
  16. Hrenovic J, Ivankovic T. Survival of Escherichia coli and Acinetobacter junii at various concentrations of sodium chloride. EurAsia J BioSci 2009;3:144–151 [CrossRef]
    [Google Scholar]
  17. Goto Y, Fink AL. Conformational states of beta-lactamase: molten-globule states at acidic and alkaline pH with high salt. Biochemistry 1989;28:945–952 [CrossRef][PubMed]
    [Google Scholar]
  18. Quan S, Wang L, Petrotchenko EV, Makepeace KA, Horowitz S et al. Super Spy variants implicate flexibility in chaperone action. Elife 2014;3:e01584 [CrossRef][PubMed]
    [Google Scholar]
  19. Qu J, Mayer C, Behrens S, Holst O, Kleinschmidt JH. The trimeric periplasmic chaperone Skp of Escherichia coli forms 1:1 complexes with outer membrane proteins via hydrophobic and electrostatic interactions. J Mol Biol 2007;374:91–105 [CrossRef][PubMed]
    [Google Scholar]
  20. Kararli TT. Comparison of the gastrointestinal anatomy, physiology, and biochemistry of humans and commonly used laboratory animals. Biopharm Drug Dispos 1995;16:351–380 [CrossRef][PubMed]
    [Google Scholar]
  21. Cabral JP. Water microbiology. Bacterial pathogens and water. Int J Environ Res Public Health 2010;7:3657–3703 [CrossRef][PubMed]
    [Google Scholar]
  22. Rozen Y, Belkin S. Survival of enteric bacteria in seawater. FEMS Microbiol Rev 2001;25:513–529 [CrossRef][PubMed]
    [Google Scholar]
  23. Sleator RD, Hill C. Bacterial osmoadaptation: the role of osmolytes in bacterial stress and virulence. FEMS Microbiol Rev 2002;26:49–71 [CrossRef][PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.000676
Loading
/content/journal/micro/10.1099/mic.0.000676
Loading

Data & Media loading...

Supplementary File 1

PDF
This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error