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  • Title: MAS dependent sensitivity of different isotopomers in selectively methyl protonated protein samples in solid state NMR.
    Author: Xue K, Sarkar R, Tosner Z, Lalli D, Motz C, Koch B, Pintacuda G, Reif B.
    Journal: J Biomol NMR; 2019 Nov; 73(10-11):625-631. PubMed ID: 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 (CH3, CH2D, CHD2) 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.
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