Characterization of Membrane Proteins in Isolated Native Cellular Membranes by Dynamic Nuclear Polarization Solid-State NMR Spectroscopy without Purification and Reconstitution

Authors

  • Dr. Tomas Jacso,

    1. Helmholtz-Zentrum München (HMGU), Deutsches Forschungszentrum für Gesundheit und Umwelt, Ingolstädter Landstrasse 1, 85764 Neuherberg (Germany)
    2. Munich Center for Integrated Protein Science (CIPS-M) at Department Chemie, Technische Universität München (TUM), Lichtenbergstrasse 4, 85747 Garching (Germany)
    3. Leibniz-Institut für Molekulare Pharmakologie (FMP), Robert-Rössle-Strasse 10, 13125 Berlin (Germany)
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  • Dr. W. Trent Franks,

    1. Leibniz-Institut für Molekulare Pharmakologie (FMP), Robert-Rössle-Strasse 10, 13125 Berlin (Germany)
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  • Dr. Honor Rose,

    1. Leibniz-Institut für Molekulare Pharmakologie (FMP), Robert-Rössle-Strasse 10, 13125 Berlin (Germany)
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  • Uwe Fink,

    1. Leibniz-Institut für Molekulare Pharmakologie (FMP), Robert-Rössle-Strasse 10, 13125 Berlin (Germany)
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  • Dr. Jana Broecker,

    1. University of Kaiserslautern, Erwin-Schrödinger-Strasse 13, 67663 Kaiserslautern (Germany)
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  • Prof. Dr. Sandro Keller,

    1. University of Kaiserslautern, Erwin-Schrödinger-Strasse 13, 67663 Kaiserslautern (Germany)
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  • Prof. Dr. Hartmut Oschkinat,

    1. Leibniz-Institut für Molekulare Pharmakologie (FMP), Robert-Rössle-Strasse 10, 13125 Berlin (Germany)
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  • Prof. Dr. Bernd Reif

    Corresponding author
    1. Helmholtz-Zentrum München (HMGU), Deutsches Forschungszentrum für Gesundheit und Umwelt, Ingolstädter Landstrasse 1, 85764 Neuherberg (Germany)
    2. Munich Center for Integrated Protein Science (CIPS-M) at Department Chemie, Technische Universität München (TUM), Lichtenbergstrasse 4, 85747 Garching (Germany)
    3. Leibniz-Institut für Molekulare Pharmakologie (FMP), Robert-Rössle-Strasse 10, 13125 Berlin (Germany)
    • Helmholtz-Zentrum München (HMGU), Deutsches Forschungszentrum für Gesundheit und Umwelt, Ingolstädter Landstrasse 1, 85764 Neuherberg (Germany)
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  • This research was supported by the Leibniz-Gemeinschaft, the Helmholtz-Gemeinschaft, and the DFG (grant number Re1435, SFB449, SFB740). W.T.F. acknowledges funding from the European Union Seventh Framework Programme (FP7/2007-2013) under the grant agreements no. 211800 (SBMPs), no. 201924 (EDICT), and no. 261863 (Bio-NMR). T.J. and B.R. are grateful to the Center for Integrated Protein Science Munich (CIPS-M) for financial support. We thank Barth van Rossum for assistance in preparing the 3D figures. We acknowledge Sascha Lange and Arne Linden for support in performing dynamic nuclear polarization experiments.

Abstract

original image

Membrane proteins in their native cellular membranes are accessible by dynamic nuclear polarization magic angle spinning solid-state NMR spectroscopy without the need of purification and reconstitution (see picture). Dynamic nuclear polarization is essential to achieve the required gain in sensitivity to observe the membrane protein of interest.

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