Role of hydrophobic interactions in yeast phosphoglycerate kinase stability.


Cold denaturation of yeast phosphoglycerate kinase (yPGK) was investigated by a combination of far UV circular dichroism (CD), steady-state and time-resolved fluorescence, and small angle X-ray scattering. It was shown that cold denaturation of yPGK cannot be accounted for by a simple two-state process and that an intermediate state can be stabilized under mild denaturing conditions. Comparison between far UV CD and fluorescence shows that in this state the protein displays a fluorescence signal corresponding mainly to exposed tryptophans, whereas its CD signal is only partially modified. Comparison with spectroscopic data obtained from a mutant missing the last 12 amino-acids (yPGK delta404) suggests that lowering the temperature mainly results in a destabilization of hydrophobic interactions between the two domains. Small angle X-ray scattering measurements give further information about this stabilized intermediate. At 4 degrees C and in the presence of 0.45 M Gdn-HCl, the main species corresponds to a protein as compact as native yPGK, whereas a significant proportion of ellipticity has been lost. Although various techniques have shown the existence of residual structures in denatured proteins, this is one example of a compact denatured state devoid of its main content in alpha helices. Study holds ProTherm entries: 7039, 7040, 7041, 7042, 7043, 7044 Extra Details: additive : EDTA(0.5 mM), phosphoglycerate kinase; cold denaturation;,hydrophobic interactions; SAXS

Submission Details

ID: Sy3agAQL

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:33 p.m.

Version: 1

Publication Details
Receveur V;Garcia P;Durand D;Vachette P;Desmadril M,Proteins (2000) Role of hydrophobic interactions in yeast phosphoglycerate kinase stability. PMID:10656268
Additional Information

Structure view and single mutant data analysis

Study data

No weblogo for data of varying length.
Colors: D E R H K S T N Q A V I L M F Y W C G P

Data Distribution

Studies with similar sequences (approximate matches)

Correlation with other assays (exact sequence matches)

Relevant PDB Entries

Structure ID Release Date Resolution Structure Title
1QPG 1996-01-04T00:00:00+0000 2.4 3-PHOSPHOGLYCERATE KINASE, MUTATION R65Q
3PGK 1982-07-15T00:00:00+0000 2.5 The structure of yeast phosphoglycerate kinase at 0.25 nm resolution

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
94.5 Phosphoglycerate kinase Q6FKY1 PGK_CANGA
100.0 Phosphoglycerate kinase P00560 PGK_YEAST