Biophysical characterization of two different stable misfolded monomeric polypeptides that are chaperone-amenable substrates.

Details

Serval ID
serval:BIB_B97FAAF79F18
Type
Article: article from journal or magazin.
Collection
Publications
Title
Biophysical characterization of two different stable misfolded monomeric polypeptides that are chaperone-amenable substrates.
Journal
Journal of Molecular Biology
Author(s)
Natalello A., Mattoo R.U., Priya S., Sharma S.K., Goloubinoff P., Doglia S.M.
ISSN
1089-8638 (Electronic)
ISSN-L
0022-2836
Publication state
Published
Issued date
2013
Volume
425
Number
7
Pages
1158-1171
Language
english
Abstract
Misfolded polypeptide monomers may be regarded as the initial species of many protein aggregation pathways, which could accordingly serve as primary targets for molecular chaperones. It is therefore of paramount importance to study the cellular mechanisms that can prevent misfolded monomers from entering the toxic aggregation pathway and moreover rehabilitate them into active proteins. Here, we produced two stable misfolded monomers of luciferase and rhodanese, which we found to be differently processed by the Hsp70 chaperone machinery and whose conformational properties were investigated by biophysical approaches. In spite of their monomeric nature, they displayed enhanced thioflavin T fluorescence, non-native β-sheets, and tertiary structures with surface-accessible hydrophobic patches, but differed in their conformational stability and aggregation propensity. Interestingly, minor structural differences between the two misfolded species could account for their markedly different behavior in chaperone-mediated unfolding/refolding assays. Indeed, only a single DnaK molecule was sufficient to unfold by direct clamping a misfolded luciferase monomer, while, by contrast, several DnaK molecules were necessary to unfold the more resistant misfolded rhodanese monomer by a combination of direct clamping and cooperative entropic pulling.
Pubmed
Web of science
Create date
03/05/2013 20:36
Last modification date
20/08/2019 16:27
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