Nearest-neighbor NMR spectroscopy: categorizing spectral peaks by their adjacent nuclei.
Nearest-neighbor NMR spectroscopy: categorizing spectral peaks by their adjacent nuclei.
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DOI：
10.1038/s41467-020-19325-4
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发表时间：
2020-11-03
影响因子：
16.6
通讯作者：
Arthanari H
中科院分区：
文献类型：
--
作者：
Behera SP;Dubey A;Chen WN;De Paula VS;Zhang M;Sgourakis NG;Bermel W;Wagner G;Coote PW;Arthanari H
甲基NMR使原子分辨率的结构和动力学的大蛋白质在溶液中的研究。然而，共振分配仍然具有挑战性。问题是联合收割机将现有的结构信息与稀疏距离约束相结合，并在排列中搜索最兼容的分配。将峰预先分类为来自异亮氨酸、亮氨酸或缬氨酸，将搜索空间减少了许多数量级。然而，这受到重叠的亮氨酸和缬氨酸频率的阻碍。相比之下，与甲基碳耦合的最近邻原子核在不同的频带中共振。在这里，我们开发了一个框架，将被动耦合共振的额外信息印在观察到的峰上。这取决于使用来自控制理论的原理，沿着沿着不同的磁化轨迹同时编排紧密间隔的谐振带。对于甲基-NMR，该方法作为标准指纹图谱（2D-HMQC）的修改实施。氨基酸类型在指纹图谱中立即显现。没有额外的弛豫损失或实验时间的增加。该方法在生物学相关蛋白质上进行了验证。使用被动的相邻共振产生新的光谱信息的想法适用于NMR光谱学中的其他背景。大型蛋白质和复合物的结构和动力学可以通过甲基NMR进行研究，但共振归属仍然具有挑战性。在这里，作者提出了一种NMR方法，该方法利用最佳控制脉冲设计，使用简单的2D HMQC实验明确区分Leu和瓦尔，并将其应用于几种蛋白质，包括Cas9，白细胞介素和人翻译起始因子eIF 4a。
Methyl-NMR enables atomic-resolution studies of structure and dynamics of large proteins in solution. However, resonance assignment remains challenging. The problem is to combine existing structural informational with sparse distance restraints and search for the most compatible assignment among the permutations. Prior classification of peaks as either from isoleucine, leucine, or valine reduces the search space by many orders of magnitude. However, this is hindered by overlapped leucine and valine frequencies. In contrast, the nearest-neighbor nuclei, coupled to the methyl carbons, resonate in distinct frequency bands. Here, we develop a framework to imprint additional information about passively coupled resonances onto the observed peaks. This depends on simultaneously orchestrating closely spaced bands of resonances along different magnetization trajectories, using principles from control theory. For methyl-NMR, the method is implemented as a modification to the standard fingerprint spectrum (the 2D-HMQC). The amino acid type is immediately apparent in the fingerprint spectrum. There is no additional relaxation loss or an increase in experimental time. The method is validated on biologically relevant proteins. The idea of generating new spectral information using passive, adjacent resonances is applicable to other contexts in NMR spectroscopy. The structure and dynamics of large proteins and complexes can be studied by methyl-NMR but resonance assignment is still challenging. Here, the authors present a NMR method that leverages optimal control pulse design to unambiguously distinguish between Leu and Val using a simple 2D HMQC experiment and they apply it to several proteins including Cas9, interleukin, and human translation initiation factor eIF4a.
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影响因子：
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