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Key stabilizing elements of protein structure identified through pressure and temperature perturbation of its hydrogen bond network

Nature Chemistry volume 4, pages 711–717 (2012)Cite this article

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Abstract

Hydrogen bonds are key constituents of biomolecular structures, and their response to external perturbations may reveal important insights about the most stable components of a structure. NMR spectroscopy can probe hydrogen bond deformations at very high resolution through hydrogen bond scalar couplings (HBCs). However, the small size of HBCs has so far prevented a comprehensive quantitative characterization of protein hydrogen bonds as a function of the basic thermodynamic parameters of pressure and temperature. Using a newly developed pressure cell, we have now mapped pressure- and temperature-dependent changes of 31 hydrogen bonds in ubiquitin by measuring HBCs with very high precision. Short-range hydrogen bonds are only moderately perturbed, but many hydrogen bonds with large sequence separations (high contact order) show greater changes. In contrast, other high-contact-order hydrogen bonds remain virtually unaffected. The specific stabilization of such topologically important connections may present a general principle with which to achieve protein stability and to preserve structural integrity during protein function.

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Figure 1: Pressure and temperature stability of proteins according to Hawley's theory.
Figure 2: High-sensitivity quantitative detection of hydrogen bonds in human ubiquitin by h3JNC′ correlation spectroscopy.
Figure 3: Pressure-induced variations of all detected hydrogen-bond correlations in ubiquitin.
Figure 4: Combined pressure and temperature dependence of representative hydrogen-bond correlations in ubiquitin.
Figure 5: Pressure and temperature derivatives of stability of h3JNC′ as quantitative measures of hydrogen-bond stability.
Figure 6: Immediate vicinity of high- and low-stability hydrogen bonds in β-sheet β3/β5.

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Acknowledgements

The authors acknowledge H.-R. Kalbitzer for introducing them to high-pressure NMR technology and R. Peterson (Daedalus Innovations) for many helpful discussions. This work was supported by SNF grant 31-132857 (to S.G.) and a stipend from the Boehringer Ingelheim Fonds (to L.N.). This article is dedicated to Anna and Joachim Seelig, celebrating their 40 years of science and life at the Biozentrum Basel.

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  1. Division of Structural Biology, Biozentrum, University of Basel, Basel, CH-4056, Switzerland

    Lydia Nisius & Stephan Grzesiek

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  1. Lydia Nisius
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L.N. and S.G. performed the experiments, analysed the data and wrote the Article.

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Correspondence to Stephan Grzesiek.

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Nisius, L., Grzesiek, S. Key stabilizing elements of protein structure identified through pressure and temperature perturbation of its hydrogen bond network. Nature Chem 4, 711–717 (2012). https://doi.org/10.1038/nchem.1396

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