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Site-resolved measurement of water-protein interactions by solution NMR

Nature Structural & Molecular Biology volume 18, pages 245–249 (2011)Cite this article

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Abstract

The interactions of biological macromolecules with water are fundamental to their structure, dynamics and function. Historically, characterization of the location and residence times of hydration waters of proteins in solution has been quite difficult. Confining proteins within the nanoscale interior of a reverse micelle slows water dynamics, allowing global protein-water interactions to be detected using nuclear magnetic resonance techniques. Complications that normally arise from hydrogen exchange and long-range dipolar coupling are overcome by the nature of the reverse micelle medium. Characterization of the hydration of ubiquitin demonstrates that encapsulation within a reverse micelle allows detection of dozens of hydration waters. Comparison of nuclear Overhauser effects obtained in the laboratory and rotating frames indicate a considerable range of hydration water dynamics is present on the protein surface. In addition, an unprecedented clustering of different hydration-dynamics classes of sites is evident.

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Figure 1: Schematic of ubiquitin in an AOT reverse micelle.
Figure 2: Detection of protein hydration by solution NMR.
Figure 3: Ubiquitin hydration dynamics map.
Figure 4: Comparison of crystallographic and solution NMR-detected hydration of human ubiquitin.

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Acknowledgements

We are grateful to K. Valentine, J. Gledhill and J. Dogan for helpful discussion and to B. Halle for comments on an early draft of this manuscript. Supported by a grant from the US National Science Foundation (MCB-0842814) and a grant from the Mathers Foundation. N.V.N. is the recipient of a US National Institutes of Health postdoctoral fellowship (GM 087099).

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Authors and Affiliations

  1. Johnson Research Foundation, University of Pennsylvania, Philadelphia, Pennsylvania, USA

    Nathaniel V Nucci, Maxim S Pometun & A Joshua Wand

  2. Department of Biochemistry and Biophysics, University of Pennsylvania, Philadelphia, Pennsylvania, USA

    Nathaniel V Nucci, Maxim S Pometun & A Joshua Wand

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  1. Nathaniel V Nucci
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  2. Maxim S Pometun
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Contributions

A.J.W. designed and initiated the study. M.S.P. carried out preliminary NMR experiments. N.V.N. optimized the NMR experiments, and collected and analyzed the data. A.J.W. and N.V.N. wrote the manuscript.

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Correspondence to A Joshua Wand.

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Competing interests

A.J.W. declares a competing financial interest as a member of Daedalus Innovations, LLC, a manufacturer of high pressure and reverse micelle NMR apparatus.

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Nucci, N., Pometun, M. & Wand, A. Site-resolved measurement of water-protein interactions by solution NMR. Nat Struct Mol Biol 18, 245–249 (2011). https://doi.org/10.1038/nsmb.1955

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