Abstract
A procedure is presented for the substantial simplification of 2D constant-time 13C−1H heteronuclear single-quantum correlation (HSQC) spectra of 13C-enriched proteins. In this approach, a single pulse sequence simultaneously records eight sub-spectra wherein the phases of the NMR signals depend on spin topology. Signals from different chemical groups are then stratified into different sub-spectra through linear combination based on Hadamard encoding of 13CHn multiplicity (n = 1, 2, and 3) and the chemical nature of neighboring 13C nuclei (aliphatic, carbonyl/carboxyl, aromatic). This results in five sets of 2D NMR spectra containing mutually exclusive signals from: (i) 13Cβ−1Hβ correlations of asparagine and aspartic acid, 13Cγ−1Hγ correlations of glutamine and glutamic acid, and 13Cα−1Hα correlations of glycine, (ii) 13Cα−1Hα correlations of all residues but glycine, and (iii) 13Cβ−1Hβ correlations of phenylalanine, tyrosine, histidine, and tryptophan, and the remaining (iv) aliphatic 13CH2 and (v) aliphatic 13CH/13CH3 resonances. As HSQC is a common element of many NMR experiments, the spectral simplification proposed in this article can be straightforwardly implemented in experiments for resonance assignment and structure determination and should be of widespread utility.
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Acknowledgements
This work was performed under the Cooperative Research Programs at the Institute for Protein Research, Osaka University (NMRCR-17-05 and NMRCR-18-05). We thank Dr. Yohei Miyanoiri (Institute for Protein Research, Osaka University) for help with NMR analysis. This work was supported by JSPS KAKENHI (Grant Number JP 19K14677) and the Uehara Memorial Foundation.
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Yoshimura, Y., Mulder, F.A.A. Sensitive and simplified: a combinatorial acquisition of five distinct 2D constant-time 13C−1H NMR protein correlation spectra. J Biomol NMR 74, 695–706 (2020). https://doi.org/10.1007/s10858-020-00341-x
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DOI: https://doi.org/10.1007/s10858-020-00341-x