Microsecond folding of the cold shock protein measured by a pressure-jump technique.


Abstract

A pressure-jump apparatus was employed in investigating the kinetics of protein unfolding and refolding. In the reaction cell, the pressure can be increased or decreased by 100-160 bar within 50-100 microseconds and then held constant. Thus, unfolding and refolding reactions in the time range from 70 microseconds to 70 s can be followed with this technique. Measurements are possible in the transition regions of thermally or denaturant-induced folding in a wide range of temperatures and solvent conditions. We used this pressure-jump method to determine the temperature dependence of the rate constants of unfolding and refolding of the cold shock protein of Bacillus subtilis and of three variants thereof with Phe --> Ala substitutions in the central beta-sheet region. For all variants, the change in heat capacity occurred in refolding between the unfolded and activated states, suggesting that the overall native-like character of the activated state of folding was not changed by the deletion of individual Phe side chains. The Phe27Ala mutation affected the rate of unfolding only; the Phe15Ala and Phe17Ala mutations changed the kinetics of both unfolding and refolding. Although the activated state of folding of the cold shock protein is overall native-like, individual side chains are still in a non-native environment. Study holds ProTherm entries: 16335, 16336, 16337, 16338, 16339, 16340, 16341, 16342, 16343, 16344, 16345, 16346 Extra Details: Thermodynamic parameters for unfolding (Native to Unfolding) cold shock protein, unfolding and refolding kinetics, pressure-jump technique

Submission Details

ID: M7q7aHzp3

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:47 p.m.

Version: 1

Publication Details
Jacob M;Holtermann G;Perl D;Reinstein J;Schindler T;Geeves MA;Schmid FX,Biochemistry (1999) Microsecond folding of the cold shock protein measured by a pressure-jump technique. PMID:10074340
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
1NMG 1996-07-11 MAJOR COLD-SHOCK PROTEIN, NMR, MINIMIZED AVERAGE STRUCTURE
2F52 2006-09-19 Solution structure of cold shock protein CspB from Bacillus subtilis in complex with heptathymidine
1NMF 1996-07-11 MAJOR COLD-SHOCK PROTEIN, NMR, 20 STRUCTURES
3PF4 2011-09-21 1.38 Crystal structure of Bs-CspB in complex with r(GUCUUUA)
3PF5 2011-09-21 1.68 Crystal structure of Bs-CspB in complex with rU6
2ES2 2006-09-05 1.78 Crystal Structure Analysis of the Bacillus Subtilis Cold Shock Protein Bs-CspB in Complex with Hexathymidine
2I5M 2007-05-22 2.3 Crystal structure of Bacillus subtilis cold shock protein CspB variant A46K S48R
1CSP 1995-05-12 2.45 CRYSTAL STRUCTURE OF THE BACILLUS SUBTILIS MAJOR COLD SHOCK PROTEIN, CSPB: A UNIVERSAL NUCLEIC-ACID BINDING DOMAIN
2I5L 2007-05-22 2.55 Crystal structure of Bacillus subtilis Cold Shock Protein variant Bs-CspB M1R/E3K/K65I
1CSQ 1995-05-12 2.7 CRYSTAL STRUCTURE OF THE BACILLUS SUBTILIS MAJOR COLD SHOCK PROTEIN, CSPB: A UNIVERSAL NUCLEIC-ACID BINDING DOMAIN

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
96.5 Cold shock protein CspB P41018 CSPB_SPOGL
92.3 Cold shock protein CspB Q816H3 CSPD_BACCR
92.3 Cold shock protein CspB Q81K90 CSPD_BACAN
100.0 Cold shock protein CspB P32081 CSPB_BACSU