Analysis of coordinated NMR chemical shifts to map allosteric regulatory networks in proteins
Analysis of coordinated NMR chemical shifts to map allosteric regulatory networks in proteins
复制标题
分析协调 NMR 化学位移以绘制蛋白质中的变构调节网络
DOI：
10.1016/j.ymeth.2022.12.002
复制
发表时间：
2023
期刊：
影响因子：
4.8
通讯作者：
Lisi, George P.
中科院分区：
文献类型：
--
作者：
Skeens, Erin;Lisi, George P.
核磁共振化学位移对局部环境的敏感性使其成为评估各种大小和结构组成的蛋白质中原子水平扰动的理想探针。生物分子的溶液和固态核磁共振波谱的最新进展利用化学转移来报告单个氨基酸之间的短期和远程偶联，以建立残基“网络”，形成变速途径的基础，通过蛋白质基质传递化学信号以诱导功能反应。一个简单的前提是，蛋白质的热力学和功能耦合区域（即活性和变容位点）对结构或动态扰动应该是相互敏感的，这使得核磁共振波谱学——蛋白质结构波动的分子分辨率的首要方法——在蛋白质变容研究中占据了前沿地位。在这里，我们详细介绍了几种核磁共振化学位移分析的关键方法，以提取蛋白质中远程化学信号的机制信息，重点是实用的方法方面和给定方法相关的情况。我们还详细介绍了将这些方法应用于特定蛋白质系统时应考虑的一些实验因素。
The exquisite sensitivity of the NMR chemical shift to local environment makes it an ideal probe to assess atomic level perturbations in proteins of all sizes and structural compositions. Recent advances in solution and solid-state NMR spectroscopy of biomolecules have leveraged the chemical shift to report on short- and long-range couplings between individual amino acids to establish “networks” of residues that form the basis of allosteric pathways that transmit chemical signals through the protein matrix to induce functional responses. The simple premise that thermodynamically and functionally coupled regions of a protein (i.e. active and allosteric sites) should be reciprocally sensitive to structural or dynamic perturbations has enabled NMR spectroscopy, the premier method for molecular resolution of protein structural fluctuations, to occupy a place at the forefront of investigations into protein allostery. Here, we detail several key methods of NMR chemical shift analysis to extract mechanistic information about long-range chemical signaling in a protein, focusing on practical methodological aspects and the circumstances under which a given approach would be relevant. We also detail some of the experimental considerations that should be made when applying these methods to specific protein systems.