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Kinetic partitioning of protein folding and aggregation

Nature Structural Biology volume 9, pages 137–143 (2002)Cite this article

Abstract

We have systematically studied the effects of 40 single point mutations on the conversion of the denatured form of the α/β protein acylphosphatase (AcP) into insoluble aggregates. All the mutations that significantly perturb the rate of aggregation are located in two regions of the protein sequence, residues 16–31 and 87–98, each of which has a relatively high hydrophobicity and propensity to form β-sheet structure. The measured changes in aggregation rate upon mutation correlate with changes in the hydrophobicity and β-sheet propensity of the regions of the protein in which the mutations are located. The two regions of the protein sequence that determine the aggregation rate are distinct from those parts of the sequence that determine the rate of protein folding. Dissection of the protein into six peptides corresponding to different regions of the sequence indicates that the kinetic partitioning between aggregation and folding can be attributed to the intrinsic conformational preferences of the denatured polypeptide chain.

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Figure 1: Rate of aggregation for wild type AcP and some representative mutational variants.
Figure 2: Change in the rate of a, aggregation and b, folding of AcP resulting from mutation at various positions.
Figure 3: β-sheet propensity and hydropathy profiles of AcP.
Figure 4: Far-UV CD spectra of peptides corresponding to various sequence regions of AcP.
Figure 5: Analysis of the changes of aggregation rate of AcP resulting from mutation.
Figure 6: Regions of AcP involved in aggregation and folding.

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Acknowledgements

We are very grateful for support from the Accademia Nazionale dei Lincei (F.C.), the Fondazione Telethon-Italia (F.C.) and the Wellcome Trust (C.M.D.). The OCMS is supported by the BBSRC, the EPSRC and the MRC. The DSB in Florence is supported by the Italian CNR and the Fondazione Telethon-Italia. The authors thank the Centro Interdip. from the University of Florence for assistance in mass spectrometry; and M. Vendruscolo, K. Plaxco, M. Karplus and A. Fersht for stimulating discussions.

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  1. Christopher M. Dobson

    Present address: Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge, CB2 1EW, UK

Authors and Affiliations

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

    Fabrizio Chiti, Niccolò Taddei, Fabiana Baroni, Cristina Capanni, Massimo Stefani & Giampietro Ramponi

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

    Fabrizio Chiti & Christopher M. Dobson

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Correspondence to Christopher M. Dobson.

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Chiti, F., Taddei, N., Baroni, F. et al. Kinetic partitioning of protein folding and aggregation. Nat Struct Mol Biol 9, 137–143 (2002). https://doi.org/10.1038/nsb752

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