Abstract：
The structural determination of biological macromolecules has been transformative for understanding biochemical mechanisms and developing therapeutics. However, the ultimate goal of characterizing how structural dynamics underpin biochemical processes has been difficult. This is largely due to significant technical challenges that hinder data collection and analysis on the native timescales of macromolecular dynamics. Single-particle cryo-EM provides a powerful platform to approach this challenge, since samples can be frozen faster than the single-turnover timescales of most biochemical reactions. In order to enable time-resolved analysis, significant innovations in the handling and preparation of cryo-EM samples have been implemented, bringing us closer to the goal of the direct observation of protein dynamics in the milliseconds to seconds range. Here, the current state of time-resolved cryo-EM is reviewed and the most promising future research directions are discussed.
摘要：
生物大分子的结构确定对于理解生化机制和开发疗法具有革命性。然而,表征结构动力学如何支撑生化过程的最终目标一直很困难。这在很大程度上是由于阻碍了对大分子动力学的天然时间尺度的数据收集和分析的重大技术挑战。单粒子低温EM提供了一个强大的平台来应对这一挑战，因为样品可以比大多数生化反应的单次周转时间更快地冷冻。为了实现时间分辨分析，在低温EM样品的处理和制备方面已经实现了重大创新，使我们更接近在毫秒到秒范围内直接观察蛋白质动力学的目标。这里,回顾了时间分辨低温EM的现状，并讨论了最有希望的未来研究方向。
