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The energy landscape of a fast-folding protein mapped by Ala→Gly Substitutions

Nature Structural Biology volume 4, pages 305–310 (1997)Cite this article

Abstract

A moderately stable protein with typical folding kinetics unfolds and refolds many times during its cellular lifetime. In monomeric λ, repressor this process is extremely rapid, with an average folded state lifetime of only 30 milliseconds. A thermostable variant of this protein (G46A/G48A) unfolds with the wild-type rate, but it folds in approximately 20 μs making it the fastest-folding protein yet observed. The effects of alanine to glycine substitutions on the folding and unfolding rate constants of the G46A/G48A variant, measured by dynamic NMR spectroscopy, indicate that the transition state is an ensemble comprised of a disperse range of conformations. This structural diversity in the transition state is consistent with the idea that folding chains are directed towards the native state by a smooth funnel-like conformational energy landscape. The kinetic data for the folding of monomeric λ repressor can be understood by merging the new energy landscape view of folding with traditional models. This hybrid model incorporates the conformational diversity of denatured and transition state ensembles, a transition state activation energy, and the importance of intrinsic helical stabilities.

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Author notes

  1. Guewha S. Huang

    Present address: Division of Structural Biology, Institute of Biomedical Sciences, Academia Sinica, Taipei, Taiwan, R.O.C

  2. Margaret A. Daugherty

    Present address: Pennsylvania State University College of Medicine, Department of Biochemistry and Molecular Biology, P.O. Box 850, Hershey, Pennsylvania, 17033, USA

Authors and Affiliations

  1. Department of Biochemistry, Duke University Medical Center, Durham, North Carolina, 27710, USA

    Randall E. Burton, Guewha S. Huang, Margaret A. Daugherty, Tiffany L. Calderone & Terrence G. Oas

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Burton, R., Huang, G., Daugherty, M. et al. The energy landscape of a fast-folding protein mapped by Ala→Gly Substitutions. Nat Struct Mol Biol 4, 305–310 (1997). https://doi.org/10.1038/nsb0497-305

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