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NMR Spectroscopic Assignment of Backbone and Side‐Chain Protons in Fully Protonated Proteins: Microcrystals,Sedimented Assemblies,and Amyloid Fibrils |
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| Authors: | Dr Jan Stanek Dr Loren B Andreas Dr Kristaps Jaudzems Dr Diane Cala Dr Daniela Lalli Andrea Bertarello Dr Tobias Schubeis Dr Inara Akopjana Dr Svetlana Kotelovica Prof Kaspars Tars Dr Andrea Pica Dr Serena Leone Prof Delia Picone Dr Zhi‐Qiang Xu Prof Nicholas E Dixon Dr Denis Martinez Mélanie Berbon Nadia El?Mammeri Dr Abdelmajid Noubhani Dr Sven Saupe Dr Birgit Habenstein Dr Antoine Loquet Dr Guido Pintacuda |
| Affiliation: | 1. Centre de RMN à Très Hauts Champs, Institut des Sciences Analytiques (UMR 5280 – CNRS, ENS Lyon, UCB Lyon 1), Université de Lyon, Villeurbanne, France;2. Biomedical Research and Study Centre, Riga, Latvia;3. Department of Chemical Sciences, University of Naples Federico II, Naples, Italy;4. School of Chemistry, University of Wollongong, NSW, Australia;5. Institute of Chemistry & Biology of Membranes & Nanoobjects (UMR 5248 CBMN – CNRS, University of Bordeaux, Bordeaux INP), All. Geoffroy Saint-Hillaire, Pessac, France;6. Institut de Biochimie et de Génétique Cellulaire (UMR 5095, CNRS –, Université de Bordeaux), Bordeaux, France |
| Abstract: | We demonstrate sensitive detection of alpha protons of fully protonated proteins by solid‐state NMR spectroscopy with 100–111 kHz magic‐angle spinning (MAS). The excellent resolution in the Cα‐Hα plane is demonstrated for 5 proteins, including microcrystals, a sedimented complex, a capsid and amyloid fibrils. A set of 3D spectra based on a Cα–Hα detection block was developed and applied for the sequence‐specific backbone and aliphatic side‐chain resonance assignment using only 500 μg of sample. These developments accelerate structural studies of biomolecular assemblies available in submilligram quantities without the need of protein deuteration. |
| Keywords: | magic-angle spinning proton detection resonance assignment solid-state NMR spectroscopy |
