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Spectral editing of alanine, serine, and threonine in uniformly labeled proteins based on frequency-selective homonuclear recoupling in solid-state NMR

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Abstract

Spectral editing is crucial to simplify the crowded solid-state NMR spectra of proteins. New techniques are introduced to edit 13C-13C correlations of uniformly labeled proteins under moderate magic-angle spinning (MAS), based on our recent frequency-selective homonuclear recoupling sequences [Zhang et al., J. Phys. Chem. Lett. 2020, 11, 8077–8083]. The signals of alanine, serine, or threonine residues are selected out by selective 13Cα-13Cβ double-quantum filtering (DQF). The 13Cα-13Cβ correlations of alanine residues are selectively established with efficiency up to ~ 1.8 times that by dipolar-assisted rotational resonance (DARR). The techniques are shown in 2D/3D NCCX experiments and applied to the uniformly 13C, 15N labeled Aquaporin Z (AqpZ) membrane protein, demonstrating their potential to simplify spectral analyses in biological solid-state NMR.

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Acknowledgements

This work was supported by grants from the National Key R&D Program of China (2016YFA0501200, 2017YFA0505400), the National Natural Science Foundation of China (21775161, 22074153, 21927801, 31627803, 31770798 and 21921004) and Chinese Academy of Sciences (YJKYYQ20190032).

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Authors and Affiliations

  1. State Key Laboratory of Magnetic Resonance and Atomic and Molecular Physics, Innovation Academy for Precision Measurement Science and Technology, National Center for Magnetic Resonance in Wuhan, Key Laboratory of Magnetic Resonance in Biological Systems, Wuhan Institute of Physics and Mathematics, Chinese Academy of Sciences, Wuhan, 430071, P. R. China

    Hang Xiao, Zhengfeng Zhang, Yongxiang Zhao & Jun Yang

  2. University of Chinese Academy of Sciences, Beijing, 100049, P. R. China

    Hang Xiao & Zhengfeng Zhang

  3. Wuhan National Laboratory for Optoelectronics, Huazhong University of Science and Technology, Wuhan, 430074, P. R. China

    Jun Yang

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  1. Hang Xiao
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Correspondence to Zhengfeng Zhang or Jun Yang.

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Xiao, H., Zhang, Z., Zhao, Y. et al. Spectral editing of alanine, serine, and threonine in uniformly labeled proteins based on frequency-selective homonuclear recoupling in solid-state NMR. J Biomol NMR 75, 193–202 (2021). https://doi.org/10.1007/s10858-021-00367-9

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