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Nonprolyl cis peptide bonds in unfolded proteins cause complex folding kinetics

Nature Structural Biology volume 8, pages 452–458 (2001)Cite this article

Abstract

Folding of tendamistat, an inhibitor of α-amylase, is a fast two-state process accompanied by two minor slow reactions, which were assigned to prolyl isomerization. In a proline-free variant, 5% of the molecules still fold slowly with a rate constant of 2.5 s−1. This reaction is caused by a slow equilibrium between two populations of unfolded molecules. The time constant for this equilibration process, its sensitivity to LiCl and its temperature dependence identify it as a cis-trans isomerization of nonprolyl peptide bonds. Although nonprolyl peptide bonds have the cis conformation populating only ∼0.15% in unfolded proteins, their large number generates a significant fraction of slow-folding molecules. This emphasizes that heterogeneous populations in an unfolded protein can induce complex folding kinetics on various time scales.

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Figure 1: Effect of proline replacements on tendamistat stability and folding.
Figure 2: Refolding kinetics of the proline-free tendamistat variant.
Figure 3: Time course of formation of the native state of proline-free tendamistat.
Figure 4: Double jump experiments to monitor the slow equilibration process in unfolded proline-free tendamistat.
Figure 5: Temperature dependence of the two folding phases.
Figure 6: Effect of LiCl on the cis-trans equilibrium of a nonprolyl peptide bond.
Figure 7: LiCl dependence of tendamistat folding.

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  1. Günter Pappenberger

    Present address: The Institute of Cancer Research, Chester Beatty Laboratories, 237 Fulham Road, London, SW3 6JB, UK

Authors and Affiliations

  1. Abteilung Biophysikalische Chemie, Biozentrum der Universität Basel, Klingelbergstrasse 70, Basel, CH-4056, Switzerland

    Günter Pappenberger & Thomas Kiefhaber

  2. Institut für Organische Chemie der Universität Frankfurt, Marie Curie Strasse 11, Frankfurt/Main, D-60439, Germany

    Hüseyin Aygün, Joachim W. Engels & Ulf Reimer

  3. Max Planck Research Unit “Enzymology of Protein Folding”, Weinbergweg 22, Halle (Saale), D-06120, Germany

    Ulf Reimer & Gunter Fischer

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Correspondence to Thomas Kiefhaber.

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Pappenberger, G., Aygün, H., Engels, J. et al. Nonprolyl cis peptide bonds in unfolded proteins cause complex folding kinetics. Nat Struct Mol Biol 8, 452–458 (2001). https://doi.org/10.1038/87624

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