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Detection of side-chain proton resonances of fully protonated biosolids in nano-litre volumes by magic angle spinning solid-state NMR

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Abstract

We present a new solid-state NMR proton-detected three-dimensional experiment dedicated to the observation of protein proton side chain resonances in nano-liter volumes. The experiment takes advantage of very fast magic angle spinning and double quantum 13C–13C transfer to establish efficient (H)CCH correlations detected on side chain protons. Our approach is demonstrated on the HET-s prion domain in its functional amyloid fibrillar form, fully protonated, with a sample amount of less than 500 µg using a MAS frequency of 70 kHz. The majority of aliphatic and aromatic side chain protons (70%) are observable, in addition to Hα resonances, in a single experiment providing a complementary approach to the established proton-detected amide-based multidimensional solid-state NMR experiments for the study and resonance assignment of biosolid samples, in particular for aromatic side chain resonances.

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Acknowledgements

We acknowledge financial support from the European Research Council (ERC) under the European Unions Horizon 2020 research and innovation programme (ERC-2015-StG GA no. 639020 to A.L.), IdEx Bordeaux (Chaire d’Installation to B.H., ANR-10-IDEX-03-02), the ANR (ANR-14-CE09-0020-01 to A.L., ANR-13-PDOC-0017-01 to B.H. and ANR-17-CE11-0035 to S.J.S) and the CNRS (IR-RMN FR3050). This work has benefited from the facilities and expertise of the Biophysical and Structural Chemistry Platform (BPCS, UMS3033).

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  1. Manoj Kumar Pandey

    Present address: Department of Chemistry, Indian Institute of Technology Ropar, Rupnagar, India

Authors and Affiliations

  1. Institute of Chemistry & Biology of Membranes & Nanoobjects, (UMR5248 CBMN), CNRS, Université Bordeaux, Institut Européen de Chimie et Biologie, 33600, Pessac, France

    James Tolchard, Mélanie Berbon, Abdelmajid Noubhani, Birgit Habenstein & Antoine Loquet

  2. JEOL RESONANCE Inc., Musashino, Akishima, Tokyo, 196-8558, Japan

    Manoj Kumar Pandey & Yusuke Nishiyama

  3. RIKEN CLST-JEOL Collaboration Center, Tsurumi, Yokohama, Kanagawa, 230-0045, Japan

    Manoj Kumar Pandey & Yusuke Nishiyama

  4. Institut de Biochimie et de Génétique Cellulaire, (UMR 5095 IBGC), CNRS, Université Bordeaux, 33077, Bordeaux, France

    Sven J Saupe

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  1. James Tolchard
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Correspondence to Yusuke Nishiyama, Birgit Habenstein or Antoine Loquet.

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Tolchard, J., Pandey, M.K., Berbon, M. et al. Detection of side-chain proton resonances of fully protonated biosolids in nano-litre volumes by magic angle spinning solid-state NMR. J Biomol NMR 70, 177–185 (2018). https://doi.org/10.1007/s10858-018-0168-3

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