Sensitive and simplified: a combinatorial acquisition of five distinct 2D constant-time 13C?1H NMR protein correlation spectra
Sensitive and simplified: a combinatorial acquisition of five distinct 2D constant-time 13C?1H NMR protein correlation spectra
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灵敏且简化：五个不同的 2D 恒定时间 13C?1H NMR 蛋白质相关光谱的组合采集
DOI：
10.1007/s10858-020-00341-x
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发表时间：
2020
影响因子：
2.7
通讯作者：
Mulder Frans A. A.
中科院分区：
文献类型：
--
作者：
Yoshimura Yuichi;Mulder Frans A. A.
本文提出了一种简化富含13 C的蛋白质的二维恒时13 C −1H异质单量子相关（HSQC）谱的方法。在该方法中，单个脉冲序列同时记录八个子谱，其中NMR信号的相位取决于自旋拓扑。来自不同化学基团的信号然后通过基于13 CHn多重性（n= 1、2和3）的Hadamard编码和相邻13 C核的化学性质（脂肪族、羰基/羧基、芳香族）的线性组合被分层到不同的子光谱中。这导致五组2D NMR光谱，其包含来自以下的互斥信号：（i）天冬酰胺和天冬氨酸的13 C β−1Hβ相关性，谷氨酰胺和谷氨酸的13 C γ−1Hγ相关性，以及甘氨酸的13 C α −1Hα相关性，（ii）除甘氨酸外所有残基的13 C α− 1Hα相关性，以及（iii）苯丙氨酸、酪氨酸、组氨酸和色氨酸的13 C β−1Hβ相关性，以及剩余的（iv）手性13 CH 2和（v）手性13 CH/13 CH 3共振。由于HSQC是许多NMR实验的共同元素，本文中提出的光谱简化可以直接在共振归属和结构测定实验中实施，应该具有广泛的实用性。
A procedure is presented for the substantial simplification of 2D constant-time13C−1H heteronuclear single-quantum correlation (HSQC) spectra of13C-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 of13CHnmultiplicity (n= 1, 2, and 3) and the chemical nature of neighboring13C nuclei (aliphatic, carbonyl/carboxyl, aromatic). This results in five sets of 2D NMR spectra containing mutually exclusive signals from: (i)13Cβ−1Hβcorrelations of asparagine and aspartic acid,13Cγ−1Hγcorrelations of glutamine and glutamic acid, and13Cα−1Hαcorrelations of glycine, (ii)13Cα−1Hαcorrelations of all residues but glycine, and (iii)13Cβ−1Hβcorrelations of phenylalanine, tyrosine, histidine, and tryptophan, and the remaining (iv) aliphatic13CH2and (v) aliphatic13CH/13CH3resonances. 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.