Contemporary hydrogen deuterium exchange mass spectrometry
Contemporary hydrogen deuterium exchange mass spectrometry
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DOI：
10.1016/j.ymeth.2018.04.023
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发表时间：
2018-07-15
期刊：
影响因子：
4.8
通讯作者：
Wilson, Derek J.
中科院分区：
文献类型：
--
作者：
Oganesyan, Irina;Lento, Cristina;Wilson, Derek J.
氢/氘交换（HDX）质谱（MS）大约在25年前作为生物化学和结构生物学的工具出现。它已成为研究蛋白质动力学，蛋白质-配体相互作用，膜蛋白和内在无序蛋白（IDP）的关键方法。在HDX标记中，蛋白质暴露于氘代溶剂（通常为D2 O）持续可变的“标记时间”，导致酰胺主链和氨基酸侧链上未受保护的不稳定质子的同位素交换。通过比较蛋白质不同区域的氘摄取水平，可以获得系统中构象和动力学变化的信息。当与MS结合时，HDX适用于探测催化和配体结合、表位作图、生物仿制药验证、候选药物筛选和膜蛋白相互作用作图等许多其他生物分析应用中的变构效应。本文通过简要介绍HDX-MS的发展，然后概述HDX理论，并最终概述了进行HDX-MS实验所涉及的方法和程序，介绍了HDX-MS。
Hydrogen/deuterium exchange (HDX) mass spectrometry (MS) emerged as a tool for biochemistry and structural biology around 25 years ago. It has since become a key approach for studying protein dynamics, protein-ligand interactions, membrane proteins and intrinsically disordered proteins (IDPs). In HDX labeling, proteins are exposed to deuterated solvent (usually D2O) for a variable 'labeling time', resulting in isotope exchange of unprotected labile protons on the amide backbone and amino acid side chains. By comparing the levels of deuterium uptake in different regions of a protein, information on conformational and dynamic changes in the system can be acquired. When coupled with MS, HDX is suitable for probing allosteric effects in catalysis and ligand binding, epitope mapping, validation of biosimilars, drug candidate screening and mapping membrane protein interactions among many other bioanalytical applications. This review introduces HDX-MS via a brief description of HDX-MS development, followed by an overview of HDX theory and ultimately an outline of methods and procedures involved in performing HDX-MS experiments.