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Protein resonance assignment at MAS frequencies approaching 100kHz: a quantitative comparison of J-coupling and dipolar-coupling-based transfer methods

Penzel, Susanne ; Smith, Albert ; Agarwal, Vipin ; Hunkeler, Andreas ; Org, Mai-Liis ; Samoson, Ago ; Böckmann, Anja ; Ernst, Matthias ; Meier, Beat

In: Journal of Biomolecular NMR, 2015, vol. 63, no. 2, p. 165-186

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    Titel
    • Protein resonance assignment at MAS frequencies approaching 100kHz: a quantitative comparison of J-coupling and dipolar-coupling-based transfer methods
    Autor
    • Penzel, Susanne. Physical Chemistry, ETH Zürich, Vladimir-Prelog-Weg 2, 8093, Zurich, Switzerland
    • Smith, Albert. Physical Chemistry, ETH Zürich, Vladimir-Prelog-Weg 2, 8093, Zurich, Switzerland
    • Agarwal, Vipin. Physical Chemistry, ETH Zürich, Vladimir-Prelog-Weg 2, 8093, Zurich, Switzerland
    • Hunkeler, Andreas. Physical Chemistry, ETH Zürich, Vladimir-Prelog-Weg 2, 8093, Zurich, Switzerland
    • Org, Mai-Liis. NMR Instituut, Tartu Teadus, Tehnomeedikum, Tallinn University of Technology, Akadeemia tee 15a, 19086, Tallinn, Estonia
    • Samoson, Ago. NMR Instituut, Tartu Teadus, Tehnomeedikum, Tallinn University of Technology, Akadeemia tee 15a, 19086, Tallinn, Estonia
    • Böckmann, Anja. Institut de Biologie et Chimie des Protéines, UMR 5086 CNRS/Université de Lyon 1, 7, passage du Vercors, 69367, Lyon, France
    • Ernst, Matthias. Physical Chemistry, ETH Zürich, Vladimir-Prelog-Weg 2, 8093, Zurich, Switzerland
    • Meier, Beat. Physical Chemistry, ETH Zürich, Vladimir-Prelog-Weg 2, 8093, Zurich, Switzerland
    Dokumententyp
    • Postprint
    Sprache
    • Englisch
    Veröffentlicht in
    • Journal of Biomolecular NMR, 2015, vol. 63, no. 2, p. 165-186. Springer Netherlands
    Andere elektronische Ausgabe
    OAI-PMH-ID
    • oai:doc.rero.ch:331554
    Summary
    We discuss the optimum experimental conditions to obtain assignment spectra for solid proteins at magic-angle spinning (MAS) frequencies around 100kHz. We present a systematic examination of the MAS dependence of the amide proton T 2′ times and a site-specific comparison of T 2′ at 93kHz versus 60kHz MAS frequency. A quantitative analysis of transfer efficiencies of building blocks, as they are used for typical 3D experiments, was performed. To do this, we compared dipolar-coupling and J-coupling based transfer steps. The building blocks were then combined into 3D experiments for sequential resonance assignment, where we evaluated signal-to-noise ratio and information content of the different 3D spectra in order to identify the best assignment strategy. Based on this comparison, six experiments were selected to optimally assign the model protein ubiquitin, solely using spectra acquired at 93kHz MAS. Within 3days of instrument time, the required spectra were recorded from which the backbone resonances have been assigned to over 96%.