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Evidence for a three-state model of protein folding from kinetic analysis of ubiquitin variants with altered core residues

Nature Structural Biology volume 3, pages 193–205 (1996)Cite this article

Abstract

To elucidate the kinetic importance of structural intermediates in single-domain proteins, we measured the effect of solution conditions and amino-acid changes at a central core residue of ubiquitin (Val 26) on the kinetics of folding and unfolding. Kinetic analysis in terms of a sequential three-state mechanism provides insight into the contribution of specific interactions within the ubiquitin core to the structural stability of the native and intermediate states. The observation that disruptive mutations and/or addition of denaturants result in an apparent two-state folding process with slower rates is explained by the destabilization of a partially folded intermediate, which is in rapid equilibrium with unfolded states. The model predicts that under sufficiently stabilizing conditions kinetic intermediates may become populated even for proteins showing apparent two-state kinetics.

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Author notes

  1. Iain D. Peters

    Present address: Hawaii Biotechnology Group, Inc., Aiea, Hawaii, 96701, USA

Authors and Affiliations

  1. Institute for Cancer Research, Fox Chase Cancer Center, 7701 Burholme Avenue, Philadelphia, Pennsylvania, 19111, USA

    Sepideh Khorasanizadeh, Iain D. Peters & Heinrich Roder

  2. Department of Chemistry, University of Pennsylvania, Philadelphia, Pennsylvania, 19104, USA

    Sepideh Khorasanizadeh

  3. Department of Biochemistry and Biophysics, University of Pennsylvania, Philadelphia, Pennsylvania, 19104, USA

    Heinrich Roder

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Khorasanizadeh, S., Peters, I. & Roder, H. Evidence for a three-state model of protein folding from kinetic analysis of ubiquitin variants with altered core residues. Nat Struct Mol Biol 3, 193–205 (1996). https://doi.org/10.1038/nsb0296-193

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