MAS dependent sensitivity of different isotopomers in selectively methyl protonated protein samples in solid state NMR.

Kai Xue, Riddhiman Sarkar, Zdenek Tosner, Daniela Lalli, Carina Motz, Benita Koch, Guido Pintacuda, Bernd Reif

Journal: Journal of biomolecular NMR 2020;73(10-11):625-631

PMID: 31515660

Abstract

Sensitivity and resolution together determine the quality of NMR spectra in biological solids. For high-resolution structure determination with solid-state NMR, proton-detection emerged as an attractive strategy in the last few years. Recent progress in probe technology has extended the range of available MAS frequencies up to above 100 kHz, enabling the detection of resolved resonances from sidechain protons, which are important reporters of structure. Here we characterise the interplay between MAS frequency in the newly available range of 70-110 kHz and proton content on the spectral quality obtainable on a 1 GHz spectrometer for methyl resonances. Variable degrees of proton densities are tested on microcrystalline samples of the α-spectrin SH3 domain with selectively protonated methyl isotopomers (CH, CHD, CHD) in a perdeuterated matrix. The experimental results are supported by simulations that allow the prediction of the sensitivity outside this experimental frequency window. Our results facilitate the selection of the appropriate labelling scheme at a given MAS rotation frequency.

Address: Helmholtz-Zentrum München (HMGU), Deutsches Forschungszentrum für Gesundheit und Umwelt, Ingolstädter Landstr. 1, 85764, Neuherberg, Germany.; Helmholtz-Zentrum München (HMGU), Deutsches Forschungszentrum für Gesundheit und Umwelt, Ingolstädter Landstr. 1, 85764, Neuherberg, Germany. [email protected].; Munich Center for Integrated Protein Science (CIPS-M) at Department Chemie, Technische Universität München (TUM), Lichtenbergstr. 4, 85747, Garching, Germany. [email protected].; Department of Chemistry, Faculty of Science, Charles University, Hlavova 8, 12842, Prague 2, Czech Republic.; Centre de Résonance Magnétique Nucléaire a Très hauts Champs (FRE 2034, CNRS, Ecole Normale Supérieure de Lyon, Université Claude Bernard Lyon 1), Université de Lyon, 5 Rue de la Doua, 69100, Villeurbanne, France.; Munich Center for Integrated Protein Science (CIPS-M) at Department Chemie, Technische Universität München (TUM), Lichtenbergstr. 4, 85747, Garching, Germany.; Dipartimento di Scienze e Innovazione Tecnologica, Università del Piemonte Orientale Amedeo Avogadro, Viale Teresa Michel, 15121, Alessandria, Italy.; Helmholtz-Zentrum München (HMGU), Deutsches Forschungszentrum für Gesundheit und Umwelt, Ingolstädter Landstr. 1, 85764, Neuherberg, Germany. [email protected].; Munich Center for Integrated Protein Science (CIPS-M) at Department Chemie, Technische Universität München (TUM), Lichtenbergstr. 4, 85747, Garching, Germany. [email protected].

DOI: 10.1007/s10858-019-00274-0

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MeSH Terms: Deuterium, Methylation, Nuclear Magnetic Resonance, Biomolecular, Protons, Sensitivity and Specificity, Spectrin, src Homology Domains