Much of our understanding of protein structure and mechanistic function has been derived from static high-resolution structures. As structural biology has continued to evolve it has become clear that high-resolution structures alone are unable to fully capture the mechanistic basis for protein structure and function in solution. Recently Hydrogen/Deuterium-exchange Mass Spectrometry (HDX-MS) has developed into a powerful and versatile tool for structural biologists that provides novel insights into protein structure and function. HDX-MS enables direct monitoring of a protein's structural fluctuations and conformational changes under native conditions in solution even as it is carrying out its functions. In this review, we focus on the use of HDX-MS to monitor these dynamic changes in proteins. We examine how HDX-MS has been applied to study protein structure and function in systems ranging from large, complex assemblies to intrinsically disordered proteins, and we discuss its use in probing conformational changes during protein folding and catalytic function. STATEMENT FOR A BROAD AUDIENCE: The biophysical and structural characterization of proteins provides novel insight into their functionalities. Protein motions, ranging from small scale local fluctuations to larger concerted structural rearrangements, often determine protein function. Hydrogen/Deuterium-exchange Mass Spectrometry (HDX-MS) has proven a powerful biophysical tool capable of probing changes in protein structure and dynamic protein motions that are often invisible to most other techniques.

中文翻译：

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使用氢/氘交换质谱法桥接蛋白质结构、动力学和功能。


我们对蛋白质结构和机械功能的大部分理解都来自静态高分辨率结构。随着结构生物学的不断发展，很明显，仅靠高分辨率结构无法完全捕获溶液中蛋白质结构和功能的机制基础。最近，氢/氘交换质谱 (HDX-MS) 已发展成为结构生物学家的一种强大且多功能的工具，为蛋白质结构和功能提供了新的见解。 HDX-MS 能够直接监测蛋白质在溶液中自然条件下的结构波动和构象变化，甚至在蛋白质发挥其功能时也是如此。在这篇综述中，我们重点关注使用 HDX-MS 监测蛋白质的这些动态变化。我们研究了如何应用 HDX-MS 来研究从大型复杂组装体到本质无序蛋白质等系统中的蛋白质结构和功能，并讨论了它在探测蛋白质折叠和催化功能过程中构象变化的用途。面向广大读者的声明：蛋白质的生物物理和结构特征提供了对其功能的新见解。蛋白质运动，从小规模的局部波动到更大范围的协调结构重排，通常决定蛋白质的功能。氢/氘交换质谱 (HDX-MS) 已被证明是一种强大的生物物理工具，能够探测大多数其他技术通常看不见的蛋白质结构和动态蛋白质运动的变化。