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Folding dynamics and mechanism of β-hairpin formation

Nature volume 390, pages 196–199 (1997)Cite this article

Abstract

Protein chains coil into α-helices and β-sheet structures. Knowing the timescales and mechanism of formation of these basic structural elements is essential for understanding how proteins fold1. For the past 40 years, α-helix formation has been extensively investigated in synthetic and natural peptides2,3,4,5, including by nanosecond kinetic studies6,7. In contrast, the mechanism of formation of β structures has not been studied experimentally. The minimal β-structure element is the β-hairpin, which is also the basic component of antiparallel β-sheets. Here we use a nanosecond laser temperature-jump apparatus to study the kinetics of folding a β-hairpin consisting of 16 amino-acid residues. Folding of the hairpin occurs in 6 µs at room temperature, which is about 30 times slower than the rate of α-helix formation6,7. We have developed a simple statistical mechanical model that provides a structural explanation for this result. Our analysis also shows that folding of a β-hairpin captures much of the basic physics of protein folding, including stabilization by hydrogen bonding and hydrophobic interactions, two-state behaviour, and a funnel-like, partially rugged energy landscape.

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Figure 1: The β-hairpin from protein G B1.
Figure 2: Thermal unfolding of the β-hairpin measured by tryptophan fluorescence (pH 7, 20 mM potassium phosphate).
Figure 3: Kinetics of β-hairpin unfolding/refolding.
Figure 4: The free-energy landscape of the β-hairpin at the midpoint of the thermal denaturation transition.

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Acknowledgements

We thank A. Szabo and P. Wolynes for helpful discussions. We also thank J.Omichinsky for providing material used in the preliminary stages of this work. V.M. was supported by a postdoctoral fellowship from the Human Frontiers Science Program Organization.

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Authors and Affiliations

  1. Laboratory of Chemical Physics, Building 5, National Institute of Diabetes, Digestive and Kidney Diseases, National Institutes of Health, Bethesda, 20892-0520, Maryland, USA

    Victor Muñoz, Victor Muñoz, Peggy A. Thompson, James Hofrichter & William A. Eaton

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  1. Victor Muñoz
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Correspondence to Victor Muñoz or William A. Eaton.

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Muñoz, V., Thompson, P., Hofrichter, J. et al. Folding dynamics and mechanism of β-hairpin formation. Nature 390, 196–199 (1997). https://doi.org/10.1038/36626

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