Background

The invention of the X-ray free-electron laser (XFEL) has provided unprecedented new opportunities for structural biology. The advantage of XFEL is an intense pulse of X-rays and a very short pulse duration (<10 fs) promising a damage-free and time-resolved crystallography approach.

Scope of review

Recent time-resolved crystallographic analyses in XFEL facility SACLA are reviewed. Specifically, metalloproteins involved in the essential reactions of bioenergy conversion including photosystem II, cytochrome c oxidase and nitric oxide reductase are described.

Major conclusions

XFEL with pump-probe techniques successfully visualized the process of the reaction and the dynamics of a protein. Since the active center of metalloproteins is very sensitive to the X-ray radiation, damage-free structures obtained by XFEL are essential to draw mechanistic conclusions. Methods and tools for sample delivery and reaction initiation are key for successful measurement of the time-resolved data.

General significance

XFEL is at the center of approaches to gain insight into complex mechanism of structural dynamics and the reactions catalyzed by biological macromolecules. Further development has been carried out to expand the application of time-resolved X-ray crystallography. This article is part of a Special Issue entitled Novel measurement techniques for visualizing ‘live’ protein molecules.

中文翻译：

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在SACLA使用X射线自由电子激光辐射对金属蛋白进行时间分辨的研究。

背景

X射线自由电子激光器（XFEL）的发明为结构生物学提供了前所未有的新机会。XFEL的优点是X射线脉冲强度高，脉冲持续时间很短（<10 fs），有望实现无损且时间分辨的晶体学方法。

审查范围

回顾了XFEL设施SACLA中最近的时间分辨晶体学分析。具体而言，描述了涉及生物能转化的基本反应的金属蛋白，包括光系统II，细胞色素c氧化酶和一氧化氮还原酶。

主要结论

使用泵探针技术的XFEL成功地可视化了反应过程和蛋白质动力学。由于金属蛋白的活性中心对X射线辐射非常敏感，因此通过XFEL获得的无损伤结构对于得出机械结论是必不可少的。样品输送和反应引发的方法和工具是成功测量时间分辨数据的关键。

一般意义

XFEL是深入了解结构动力学复杂机制以及生物大分子催化的反应的方法的中心。已经进行了进一步的开发以扩展时间分辨X射线晶体学的应用。本文是名为“用于可视化“活”蛋白分子的新型测量技术”的特刊的一部分。