Department of Chemistry, York University, Toronto, Ontario M3J 1P3, Canada.
Chem Rev. 2022 Apr 27;122(8):7624-7646. doi: 10.1021/acs.chemrev.1c00222. Epub 2021 Jul 29.
Life at the molecular level is a dynamic world, where the key players─proteins, oligonucleotides, lipids, and carbohydrates─are in a perpetual state of structural flux, shifting rapidly between local minima on their conformational free energy landscapes. The techniques of classical structural biology, X-ray crystallography, structural NMR, and cryo-electron microscopy (cryo-EM), while capable of extraordinary structural resolution, are innately ill-suited to characterize biomolecules in their dynamically active states. Subsecond time-resolved mass spectrometry (MS) provides a unique window into the dynamic world of biological macromolecules, offering the capacity to directly monitor biochemical processes and conformational shifts with a structural dimension provided by the electrospray charge-state distribution, ion mobility, covalent labeling, or hydrogen-deuterium exchange. Over the past two decades, this suite of techniques has provided important insights into the inherently dynamic processes that drive function and pathogenesis in biological macromolecules, including (mis)folding, complexation, aggregation, ligand binding, and enzyme catalysis, among others. This Review provides a comprehensive account of subsecond time-resolved MS and the advances it has enabled in dynamic structural biology, with an emphasis on insights into the dynamic drivers of protein function.
生命在分子水平上是一个动态的世界，其中关键的参与者——蛋白质、寡核苷酸、脂类和碳水化合物——处于结构流的持续状态，在构象自由能景观的局部最小值之间快速移动。经典结构生物学技术，如 X 射线晶体学、结构 NMR 和冷冻电子显微镜（cryo-EM），虽然能够达到非凡的结构分辨率，但天生不适合描述处于动态活跃状态的生物分子。亚秒级时间分辨质谱（MS）为生物大分子的动态世界提供了一个独特的窗口，能够直接监测生化过程和构象变化，并通过电喷雾荷态分布、离子迁移率、共价标记或氘氢交换提供结构维度。在过去的二十年中，这一系列技术为驱动生物大分子功能和发病机制的固有动态过程提供了重要的见解，包括（错误）折叠、络合、聚集、配体结合和酶催化等。本综述全面介绍了亚秒级时间分辨 MS 及其在动态结构生物学中的应用进展，重点介绍了对蛋白质功能的动态驱动因素的深入了解。
