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.

中文翻译：

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现代氢氘交换质谱

大约 25 年前，氢/氘交换 (HDX) 质谱 (MS) 成为生物化学和结构生物学的一种工具。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质 (IDP) 的关键方法。在 HDX 标记中，蛋白质暴露于氘化溶剂（通常是 D2O）中一段可变的“标记时间”，导致酰胺骨架和氨基酸侧链上未受保护的不稳定质子发生同位素交换。通过比较蛋白质不同区域的氘摄取水平，可以获得有关系统构象和动态变化的信息。当与 MS 结合使用时，HDX 适用于探测催化和配体结合、表位作图、生物仿制药验证中的变构效应，许多其他生物分析应用中的候选药物筛选和膜蛋白相互作用图谱。本综述通过对 HDX-MS 发展的简要描述介绍了 HDX-MS，然后是 HDX 理论的概述，最后是执行 HDX-MS 实验所涉及的方法和程序的概述。