A Novel Method for Assessing the Chaperone Activity of Proteins

PLoS One. 2016 Aug 26;11(8):e0161970. doi: 10.1371/journal.pone.0161970. eCollection 2016.

Abstract

Protein chaperones are molecular machines which function both during homeostasis and stress conditions in all living organisms. Depending on their specific function, molecular chaperones are involved in a plethora of cellular processes by playing key roles in nascent protein chain folding, transport and quality control. Among stress protein families-molecules expressed during adverse conditions, infection, and diseases-chaperones are highly abundant. Their molecular functions range from stabilizing stress-susceptible molecules and membranes to assisting the refolding of stress-damaged proteins, thereby acting as protective barriers against cellular damage. Here we propose a novel technique to test and measure the capability for protective activity of known and putative chaperones in a semi-high throughput manner on a plate reader. The current state of the art does not allow the in vitro measurements of chaperone activity in a highly parallel manner with high accuracy or high reproducibility, thus we believe that the method we report will be of significant benefit in this direction. The use of this method may lead to a considerable increase in the number of experimentally verified proteins with such functions, and may also allow the dissection of their molecular mechanism for a better understanding of their function.

MeSH terms

  • Alcohol Dehydrogenase / chemistry
  • Alcohol Dehydrogenase / metabolism
  • Citrate (si)-Synthase / chemistry
  • Citrate (si)-Synthase / metabolism
  • L-Lactate Dehydrogenase / chemistry
  • L-Lactate Dehydrogenase / metabolism
  • Luciferases / chemistry
  • Luciferases / metabolism
  • Molecular Chaperones / chemistry
  • Molecular Chaperones / metabolism*
  • Protein Array Analysis / methods*
  • Protein Binding
  • Protein Folding
  • Proteins / chemistry
  • Proteins / metabolism*

Substances

  • Molecular Chaperones
  • Proteins
  • Alcohol Dehydrogenase
  • L-Lactate Dehydrogenase
  • Luciferases
  • Citrate (si)-Synthase

Grants and funding

This work was supported by the Odysseus grant G.0029.12 from the Fonds Wetenschappelijk Onderzoek (FWO, http://www.fwo.be) to PT, and DK is supported by a FWO postdoctoral fellowship 1.2.734.13. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.