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Characterization of different water pools in solid-state NMR protein samples

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Abstract

We observed and characterized two distinct signals originating from different pools of water protons in solid-state NMR protein samples, namely from crystal water which exchanges polarization with the protein (on the NMR timescale) and is located in the protein-rich fraction at the periphery of the magic-angle spinning (MAS) sample container, and supernatant water located close to the axis of the sample container. The polarization transfer between the water and the protein can be probed by two-dimensional exchange spectroscopy, and we show that the supernatant water does not interact with protein on the timescale of the experiments. The two water pools have different spectroscopic properties, including resonance frequency, longitudinal, transverse and rotating frame relaxation times. The supernatant water can be removed almost completely physically or can be frozen selectively. Both measures lead to an enhancement of the quality factor of the probe circuit, accompanied by an improvement of the experimental signal/noise, and greatly simplify solvent-suppression by substantially reducing the water signal. We also present a tool, which allows filling solid-state NMR sample containers in a more efficient manner, greatly reducing the amount of supernatant water and maximizing signal/noise.

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Acknowledgement

We thank Michel Juy for the picture of the Crh proteins arranged in the unit cell. This work was funded in part by CNRS, the French Research Ministry (ANR JCJC JC05_44957, ANR PCV 07 PROTEIN MOTION), the Swiss National Science Foundation (SNF) and the ETH Zurich.

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Authors and Affiliations

  1. Institut de Biologie et Chimie des Protéines, Université de Lyon, UMR 5086 CNRS/UCB-Lyon 1, 7 passage du Vercors, 69367, Lyon, France

    Anja Böckmann, Carole Gardiennet & Antoine Loquet

  2. Physical Chemistry, ETH Zürich, Wolfgang-Pauli-Strasse 10, 8093, Zurich, Switzerland

    René Verel, Andreas Hunkeler & Beat H. Meier

  3. Université de Lyon, CNRS/ENS Lyon/UCB-Lyon 1, Centre RMN à Très Hauts Champs, 5 rue de la Doua, 69100, Villeurbanne, France

    Guido Pintacuda, Lyndon Emsley & Anne Lesage

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  1. Anja Böckmann
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  2. Carole Gardiennet
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  4. Andreas Hunkeler
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  5. Antoine Loquet
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  8. Beat H. Meier
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  9. Anne Lesage
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Correspondence to Anja Böckmann or Anne Lesage.

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Böckmann, A., Gardiennet, C., Verel, R. et al. Characterization of different water pools in solid-state NMR protein samples. J Biomol NMR 45, 319–327 (2009). https://doi.org/10.1007/s10858-009-9374-3

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  • DOI: https://doi.org/10.1007/s10858-009-9374-3

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