Please use this identifier to cite or link to this item:
Type: Artigo
Title: Remodeling of the folding free energy landscape of staphylococcal nuclease by cavity-creating mutations
Author: Roche, Julien
Dellarole, Mariano
Caro, Jose A.
Guca, Ewelina
Norberto, Douglas R.
Yang, Yinshan
Garcia, Angel E.
Roumestand, Christian
Garcia-Moreno, Bertrand
Royer, Catherine A.
Abstract: The folding of staphylococcal nuclease (SNase) is known to proceed via :a major intermediate in which the central OB subdomain,is folded and the C-terminal helical subdomain is disordered To identify the structural and energetic determinants of this folding free energy landscape, we have examined in detail;;using high pressure NMR, the consequences of cavity creating mutations in each of the two sub domains of an ultrastable. SNase,. Delta+PHS. The stabilizing mutations of Delta+PHS-enhanced,the population of the major folding intermediate Cavity creation in two different regions of the Delta+PHS reference protein, despite equivalent effects on global stability, had very distinct consequences on the complexity of the folding free energy landscape. The substitution in the C-terminal helix of Delta+PHS slightly suppressed the major intermediate and promoted an additional excited state involving disorder in the N-terminus, but otherwise decreased landscape heterogeneity with respect to the Delta+PHS background protein. The 192A substitution, located in the hydrophobic OB-fold core, had a much more profound effect, resulting in a significant increase in the number. of intermediate states and implicating the entire protein structure. Denaturant (GuHCl) had very subtle and specific effects on the landscape, suppressing senile states and favoring others, depending upon the mutational context These results demonstrate that disrupting interactions in a region of the protein with highly cooperative, unfrustrated folding has Very Profound effects on the roughness of the folding landscape, : whereas the effects are less pronounced for an energetically equivalent:substitution:in an already frustrated region
Subject: Energia livre
Country: Estados Unidos
Editor: American Chemical Society
Rights: Fechado
Identifier DOI: 10.1021/bi301071z
Date Issue: 2012
Appears in Collections:IB - Artigos e Outros Documentos

Files in This Item:
File Description SizeFormat 
000311522000010.pdf1.83 MBAdobe PDFView/Open

Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.