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


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 UniProtKB Entries

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