Document Detail


Mechanism of pressure-induced thermostabilization of proteins: studies of glutamate dehydrogenases from the hyperthermophile Thermococcus litoralis.
MedLine Citation:
PMID:  11514665     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
In this study, we investigated the effect of pressure on protein structure and stability at high temperature. Thermoinactivation experiments at 5 and 500 atm were performed using the wild-type (WT) enzyme and two single mutants (D167T and T138E) of the glutamate dehydrogenase (GDH) from the hyperthermophile Thermococcus litoralis. All three GDHs were stabilized, although to different degrees, by the application of 500 atm. Interestingly, the degree of pressure stabilization correlated with GDH stability as well as the magnitude of electrostatic repulsion created by residues at positions 138 and 167. Thermoinactivation experiments also were performed in the presence of trehalose. Addition of the sugar stabilized all three GDHs; the degree of sugar-induced thermostabilization followed the same order as pressure stabilization. Previous studies suggested a mechanism whereby the enzyme adopts a more compact and rigid structure and volume fluctuations away from the native state are diminished under pressure. The present results on the three GDHs allowed us to further confirm and refine the proposed mechanism for pressure-induced thermostabilization. In particular, we propose that pressure stabilizes against thermoinactivation by shifting the equilibrium between conformational substates of the GDH hexamer, thus inhibiting irreversible aggregation.
Authors:
M M Sun; R Caillot; G Mak; F T Robb; D S Clark
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S.    
Journal Detail:
Title:  Protein science : a publication of the Protein Society     Volume:  10     ISSN:  0961-8368     ISO Abbreviation:  Protein Sci.     Publication Date:  2001 Sep 
Date Detail:
Created Date:  2001-08-21     Completed Date:  2001-10-11     Revised Date:  2013-06-09    
Medline Journal Info:
Nlm Unique ID:  9211750     Medline TA:  Protein Sci     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1750-7     Citation Subset:  IM    
Affiliation:
Department of Chemical Engineering, University of California, Berkeley, California 94720, USA.
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MeSH Terms
Descriptor/Qualifier:
Enzyme Stability
Glutamate Dehydrogenase / chemistry*,  metabolism*
Hot Temperature*
Kinetics
Models, Molecular
Pressure
Protein Structure, Quaternary
Thermococcus / enzymology*
Trehalose / metabolism
Chemical
Reg. No./Substance:
99-20-7/Trehalose; EC 1.4.1.2/Glutamate Dehydrogenase
Comments/Corrections

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