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Evidence concerning rate-limiting steps in protein folding from the effects of trifluoroethanol

Nature Structural Biology volume 7, pages 58–61 (2000)Cite this article

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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Figure 1: Natural logarithm of the refolding rate constants (ln kobs) in s-1 as a function of TFE concentration for a, cAcP; b, FNIII; c, Fyn-SH3; d, BLA.
Figure 2: Correlations between a, the extent of acceleration of folding by TFE (m‡ac R-1 T-1) and the number of local backbone hydrogen bonds (NLH), in the native state, and b, between the extent of deceleration of folding by TFE (m‡dec R-1 T-1) and by GdnHCl (m‡GdnHCl R-1 T-1).
Figure 3: Schematic representations of three different scenarios for a protein folding process.
Figure 4: Correlations between the extent of deceleration (m‡dec R-1 T-1) and a, ΔASAN->U, b, ΔASAnonpolar or c, ΔASApolar.

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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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Author notes

  1. J. Iñaki Guijarro

    Present address: Laboratoire de RMN, Département SIDA-Rétrovirus, Institut Pasteur 28, rue du Dr. Roux, 75724, Paris, Cedex 15, France

  2. Mikio Kataoka

    Present address: Laboratory of Bioenergetics and Biophysics, Graduate School of Materials Science, Nara Institute of Science and Technology, Ikoma, Nara, 630-0101, Japan

Authors and Affiliations

  1. Oxford Centre for Molecular Sciences, New Chemistry Laboratory,University of Oxford, South Parks Road, Oxford, OX1 3QT, UK

    Daizo Hamada, Fabrizio Chiti, J. Iñaki Guijarro & Christopher M. Dobson

  2. Department of Earth and Space Science, Graduate School of Science, Osaka University, Machikaneyama, Toyonaka, Osaka, 560, Japan

    Mikio Kataoka

  3. Dipartimento di Scienze Biochimiche, Università degli Studi di Firenze, Viale Morgagni 50, Firenze, 50134 , Italy

    Niccoló Taddei

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  1. Daizo Hamada
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Correspondence to Christopher M. Dobson.

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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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