Jacob, Marie-Andree and Holtermann, Georg and Perl, D. and Reinstein, J. and Schindler, T. and Geeves, Michael A. and Schmid, F.X. (1999) Microsecond folding of the cold shock protein measured by a pressure-jump technique. Biochemistry, 38 (10). pp. 2882-2891. ISSN 0006-2960. (doi:https://doi.org/10.1021/bi982487i) (The full text of this publication is not currently available from this repository. You may be able to access a copy if URLs are provided)
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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 mu s and then held constant. Thus, unfolding and refolding reactions in the time range from 70 mu s 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.
|Divisions:||Faculties > Sciences > School of Biosciences|
|Depositing User:||I.T. Ekpo|
|Date Deposited:||18 Apr 2009 21:28 UTC|
|Last Modified:||21 May 2014 07:41 UTC|
|Resource URI:||https://kar.kent.ac.uk/id/eprint/16869 (The current URI for this page, for reference purposes)|