Triple resonance three-dimensional protein NMR: before it became a black box.
Triple resonance three-dimensional protein NMR: before it became a black box.
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DOI：
10.1016/j.jmr.2011.08.003
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发表时间：
2011-12-01
期刊：
影响因子：
--
通讯作者：
Bax, Ad
中科院分区：
文献类型：
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作者：
Bax, Ad
三维三重共振实验已成为几乎所有蛋白质溶液 NMR 研究不可或缺的一部分。该方法依赖于 (13)C 和 (15)N 蛋白质的均匀同位素富集，并通过大 (1)J(NH)、(1)J(CH)(、1)J(CC) 和 (1)J(CN) 耦合建立原子核之间的标量连接通路。磁化转移过程是通过多个高效的单键磁化转移步骤发生的，而不是通过较小且可变的 (3)J(HH) 耦合的单个步骤。单键耦合相对较大的尺寸和良好的均匀性允许设计有效的磁化转移方案，该方案在给定蛋白质上有效均匀，几乎独立于构象。尽管概念上很简单，但蛋白质三维三重共振实验的实际实施最初提出了严峻的挑战。本文对这项工作的一些历史背景、遇到的问题及其解决方案以及它们演变为当今标准实验库提供了个人视角。
Three-dimensional triple resonance experiments have become an integral part of virtually every solution NMR study of proteins. The approach relies on uniform isotopic enrichment of proteins with (13)C and (15)N, and establishes the scalar connectivity pathway between nuclei through the large (1)J(NH), (1)J(CH)(, 1)J(CC), and (1)J(CN) couplings. The magnetization transfer process takes place through multiple, efficient one-bond magnetization transfer steps, rather than a single step through the smaller and variable (3)J(HH) couplings. The relatively large size and good uniformity of the one-bond couplings allowed the design of efficient magnetization transfer schemes that are effectively uniform across a given protein, nearly independent of conformation. Although conceptually straightforward, practical implementation of three-dimensional triple resonance experiments on proteins originally posed serious challenges. This account provides a personal perspective on some of the historical background to this work, the problems encountered as well as their solutions, and their evolution into today's standard arsenal of experiments.