NMR Spectroscopy of Soluble Protein Complexes at One Mega-Dalton and Beyond

Authors

  • Dr. Andi Mainz,

    1. Munich Center for Integrated Protein Science (CIPS-M) at the Department Chemie, Technische Universität München (TUM), Lichtenbergstrasse 4, 85747 Garching (Germany)
    2. Helmholtz-Zentrum München (HMGU), Deutsches Forschungszentrum für Gesundheit und Umwelt, Neuherberg (Germany)
    3. Leibniz-Institut für Molekulare Pharmakologie (FMP), Berlin (Germany)
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  • Prof. Dr. Tomasz L. Religa,

    1. University of Toronto, Department of Medical Genetics and Microbiology (Canada)
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  • Dr. Remco Sprangers,

    1. Max Planck-Institute for Developmental Biology, Tübingen (Germany)
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  • Dr. Rasmus Linser ,

    1. Harvard Medical School, Department of Biological Chemistry and Molecular Pharmacology, Boston (USA)
    2. School of Chemistry, UNSW, Sydney (Australia)
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  • Prof. Dr. Lewis E. Kay,

    1. University of Toronto, Department of Medical Genetics and Microbiology (Canada)
    2. Program in Molecular Structure and Function, Hospital for Sick Children, Toronto (Canada)
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  • Prof. Dr. Bernd Reif

    Corresponding author
    1. Munich Center for Integrated Protein Science (CIPS-M) at the Department Chemie, Technische Universität München (TUM), Lichtenbergstrasse 4, 85747 Garching (Germany)
    2. Helmholtz-Zentrum München (HMGU), Deutsches Forschungszentrum für Gesundheit und Umwelt, Neuherberg (Germany)
    3. Leibniz-Institut für Molekulare Pharmakologie (FMP), Berlin (Germany)
    • Munich Center for Integrated Protein Science (CIPS-M) at the Department Chemie, Technische Universität München (TUM), Lichtenbergstrasse 4, 85747 Garching (Germany)
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  • This work was performed in the framework of SFB-1035/Project-B07 (German Research Foundation, DFG). We acknowledge support by the Leibniz-Gemeinschaft, the Helmholtz-Gemeinschaft, and the Deutsche Forschungsgemeinschaft (Grants Re1435). We are also grateful to the Center for Integrated Protein Science Munich (CIPS-M) for financial support (A.M., B.R.). This work was further supported by the Max Planck Society (R.S.) and by a grant from the Canadian Institutes of Health Research (CIHR) (T.L.R., L.E.K.). L.E.K. holds a Canada Research Chair in Biochemistry. R.L. was supported by an Australian Research Council Discovery Early Career Research Award.

Abstract

original image

Bigger is better: Sequential backbone assignments are obtained by NMR spectroscopy for a 1 MDa proteasome complex. The method relies on immobilization of a soluble protein complex by magic-angle spinning. Deuteration and proton detection of exchangeable sites and paramagnetic relaxation enhancement enables exploration of structural and dynamic properties of supramolecular assemblies at atomic resolution.

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