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Acceleration of the folding of acylphosphatase by stabilization of local secondary structure

Nature Structural Biology volume 6, pages 380–387 (1999)Cite this article

Abstract

The addition of trifluoroethanol or hexafluoroisopropanol converts the apparent two-state folding of acylphosphatase, a small α/β protein, into a multistate mechanism where secondary structure accumulates significantly in the denatured state before folding to the native state. This results in a marked acceleration of folding as revealed by following the intrinsic fluorescence and circular dichroism changes upon folding. The folding rate is at a maximum when the secondary-structure content of the denatured state corresponds to that of the native state, while further stabilization of secondary structure decreases the folding rate. These findings indicate that stabilization of intermediate structure can either enhance or retard folding depending on its nature and content of native-like interactions.

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Figure 1: Equilibrium conformational changes induced by fluoroalcohols.
Figure 2: TFE and HFIP concentration dependence of the natural logarithm of the observed folding rate constant for muscle (▴,▵) and CT (●,○) AcP.
Figure 3: Folding rate constant of CT AcP, expressed as its natural logarithm, versus urea concentration.
Figure 4: Folding of CT AcP monitored by 222 nm far-UV CD.
Figure 5: Relationship between secondary-structure content of the denatured state and folding rate.

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Acknowledgements

We are grateful to L. Serrano and M. Buck for useful discussions. F.C. was supported by a grant from the European Community. D.H. was supported by JSPS Postdoctoral Fellowships for Research Abroad. This is a contribution from the Oxford Centre for Molecular Sciences, which is funded by BBSRC, EPSRC and MRC. The work has also been supported by funds from the Italian CNR (Target Project Biotechnology), from MURST (Project Structural Biology) and from the European Community (Biotechnology Unit). The research of C.M.D. is supported in part by an International Research Scholars award from the Howard Hughes Medical Institute and by The Wellcome Trust.

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Affiliations

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

    Fabrizio Chiti, Paul Webster, Daizo Hamada & Christopher M. Dobson

  2. Dipartimento di Scienze Biochimiche, Universita' degli Studi di Firenze, Viale Morgagni 50, Firenze, 50134, Italy

    Niccolo' Taddei, Tania Fiaschi & Giampietro Ramponi

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  1. Fabrizio Chiti
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Correspondence to Christopher M. Dobson.

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Chiti, F., Taddei, N., Webster, P. et al. Acceleration of the folding of acylphosphatase by stabilization of local secondary structure. Nat Struct Mol Biol 6, 380–387 (1999). https://doi.org/10.1038/7616

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