Abstract
The refolding kinetics of 13 proteins have been studied in the presence of 2,2,2-trifluoroethanol (TFE). Low concentrations of TFE increased the folding rates of all the proteins, whereas higher concentrations have the opposite effect. The extent of deceleration of folding correlates closely with similar effects of guanidine hydrochloride and can be related to the burial of accessible surface area during folding. For those proteins folding in a two-state manner, the extent of acceleration of folding correlates closely with the number of local backbone hydrogen bonds in the native structure. For those proteins that fold in a multistate manner, however, the extent of acceleration is much smaller than that predicted from the data for two-state proteins. These results support the concept that for two-state proteins the search for native-like contacts is a key aspect of the folding reaction, whereas the rate-determining steps for folding of multistate proteins are associated with the reorganization of stable structure within a collapsed state or with the search for native-like interactions within less structured regions.
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Acknowledgements
We are very grateful to G. Ramponi, J. Clarke, Y. Goto, B.B. Kragelund, F.M. Poulsen, J. Bright, I.D. Campbell and J. Zurdo for gifts of proteins. We also acknowledge M. Buck, B.B. Kragelund, S.E. Jackson and K.W. Plaxco for valuable discussions. The Oxford Centre for Molecular Sciences is supported by the United Kingdom BBSRC, EPSRC and MRC. The Dipartimento di Scienze Biochimiche, Università di Firenze, is supported by MURST (fondi ex 40% - Folding e Misfolding di Proteine). A JSPS Postdoctoral Fellowships for Research Abroad supports D.H. This work was also supported in part by an International Research Scholars award from the Howard Hughes Medical Institute and by a programme grant from the Wellcome Trust (C.M.D.), the Grants-in-Aid for Scientific Research from the Ministry of Education, Science and Culture of Japan (M.K.) and the European Community (C.M.D., F.C. and J.I.G.).
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Hamada, D., Chiti, F., Guijarro, J. et al. Evidence concerning rate-limiting steps in protein folding from the effects of trifluoroethanol. Nat Struct Mol Biol 7, 58–61 (2000). https://doi.org/10.1038/71259
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DOI: https://doi.org/10.1038/71259
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