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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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).
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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