A guide to time-resolved structural analysis of light-activated proteins.
A guide to time-resolved structural analysis of light-activated proteins.
复制标题
光激活蛋白质的时间分辨结构分析指南。
DOI：
10.1111/febs.15880
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发表时间：
2022
期刊：
影响因子：
--
通讯作者：
Poddar H
中科院分区：
文献类型：
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作者：
Poddar H
蛋白质结构的动态变化对蛋白质功能至关重要，并且发生在飞秒到秒的时间尺度上。X射线自由电子激光以前所未有的时间和空间分辨率促进了蛋白质结构动力学的研究。光活化蛋白是时间分辨结构研究的有吸引力的目标，因为反应化学和相关的蛋白质结构变化可以由短激光脉冲触发。具有不同光吸收中心的蛋白质已经进化到可以探测光并利用光子能量来带来下游的化学和生物输出反应。在光吸收之后，快速的化学/小尺度结构变化通常发生在发色团周围。这些局部变化之后是更大的结构变化，在整个光感受器/光催化剂中传播，从而实现所需的化学和/或生物输出反应。时间分辨系列飞秒晶体学（SFX）和溶液散射技术能够在以前无法到达的时间尺度上直接可视化光激活蛋白的早期化学变化，而散射技术则可以获得与更全局结构变化相关的更慢的时间尺度。在这里，我们回顾了时间分辨SFX和溶液散射技术的进展如何揭示了光化学及其与输出响应的耦合机制。我们还展望了这些时间分辨结构方法如何影响其他尚未通过这些方法研究的光感受器/光酶。
Dynamical changes in protein structures are essential for protein function and occur over femtoseconds to seconds timescales. X‐ray free electron lasers have facilitated investigations of structural dynamics in proteins with unprecedented temporal and spatial resolution. Light‐activated proteins are attractive targets for time‐resolved structural studies, as the reaction chemistry and associated protein structural changes can be triggered by short laser pulses. Proteins with different light‐absorbing centres have evolved to detect light and harness photon energy to bring about downstream chemical and biological output responses. Following light absorption, rapid chemical/small‐scale structural changes are typically localised around the chromophore. These localised changes are followed by larger structural changes propagated throughout the photoreceptor/photocatalyst that enables the desired chemical and/or biological output response. Time‐resolved serial femtosecond crystallography (SFX) and solution scattering techniques enable direct visualisation of early chemical change in light‐activated proteins on timescales previously inaccessible, whereas scattering gives access to slower timescales associated with more global structural change. Here, we review how advances in time‐resolved SFX and solution scattering techniques have uncovered mechanisms of photochemistry and its coupling to output responses. We also provide a prospective on how these time‐resolved structural approaches might impact on other photoreceptors/photoenzymes that have not yet been studied by these methods.