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Site-specific protein methyl deuterium quadrupolar patterns by proton-detected 3D 2H–13C–1H MAS NMR spectroscopy

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Abstract

Determination of protein structure and dynamics is key to understand the mechanism of protein action. Perdeuterated proteins have been used to obtain high resolution/sensitivty NMR experiments via proton-detection. These methods utilizes 1H, 13C and 15N nuclei for chemical shift dispersion or relaxation probes, despite the existing abundant deuterons. However, a high-sensitivity NMR method to utilize deuterons and e.g. determine site-specific deuterium quadrupolar pattern information has been lacking due to technical difficulties associated with deuterium’s large quadrupolar couplings. Here, we present a novel deuterium-excited and proton-detected three-dimensional 2H–13C–1H MAS NMR experiment to utilize deuterons and to obtain site-specific methyl 2H quadrupolar patterns on detuterated proteins for the first time. A high-resolution fingerprint 1H–15N HSQC-spectrum is correlated with the anisotropic deuterium quadrupolar tensor in the third dimension. Results from a model perdeuterated protein has been shown.

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Authors and Affiliations

  1. Radboud University, Magnetic Resonance Research Center, Institute for Molecules and Materials, Nijmegen, The Netherlands

    Ümit Akbey

  2. Department of Structural Biology, University of Pittsburgh School of Medicine, 3501 Fifth Ave., Pittsburgh, PA, USA

    Ümit Akbey

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  1. Ümit Akbey
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Akbey, Ü. Site-specific protein methyl deuterium quadrupolar patterns by proton-detected 3D 2H–13C–1H MAS NMR spectroscopy. J Biomol NMR 76, 23–28 (2022). https://doi.org/10.1007/s10858-021-00388-4

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