Serial femtosecond crystallography at the SACLA: breakthrough to dynamic structural biology
Serial femtosecond crystallography at the SACLA: breakthrough to dynamic structural biology
复制标题
SACLA 的系列飞秒晶体学：动态结构生物学的突破
DOI：
10.1007/s12551-017-0344-9
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发表时间：
2017
影响因子：
--
通讯作者：
Iwata So
中科院分区：
文献类型：
--
作者：
Mizohata Eiichi;Nakane Takanori;Fukuda Yohta;Nango Eriko;Iwata So
X射线结晶学将世界视觉化到原子水平。它被用作观察生物大分子三维结构的最强大的技术，并开创了结构生物学的先河。要确定高分辨率的晶体结构，传统上需要制备大晶体(>；约200μm)。后来，产生强大X射线的同步辐射设备，如Spring8号，被建造起来。它们使用户即使使用较小的晶体(约200-50μm)也能获得高质量的X射线衍射图像。近年来，X射线自由电子激光器(XFELs)的发展是结构生物学领域最重要的技术创新之一。日本的Spring-8 Angstrom紧凑型自由电子激光器(SACLA)通过将电子加速到相对论速度并引导它们通过真空中的短周期波荡器来产生XFEL光束。自2012年开始用户操作以来，我们一直参与SACLA使用XFEL的系列飞秒晶体(SFX)测量系统的开发。SACLA产生的X射线比Spring8号的亮度高10亿倍。极亮的XFEL脉冲使微晶体(约50-1μm)能够进行数据采集。尽管存在许多分子分析技术，但SFX是唯一一种可以在室温下以原子分辨率和快速时间分辨率可视化生物大分子无辐射损伤结构的技术。在这里，我们回顾了SACLA-SFX项目在过去5年中取得的成就。我们特别关注：(1)SFX的测量系统；(2)SFX的实验定相；(3)基于无损伤的室温结构的酶化学；(4)时间分辨SFX拍摄的分子电影。
X-ray crystallography visualizes the world at the atomic level. It has been used as the most powerful technique for observing the three-dimensional structures of biological macromolecules and has pioneered structural biology. To determine a crystal structure with high resolution, it was traditionally required to prepare large crystals (> 200 μm). Later, synchrotron radiation facilities, such as SPring-8, that produce powerful X-rays were built. They enabled users to obtain good quality X-ray diffraction images even with smaller crystals (ca. 200–50 μm). In recent years, one of the most important technological innovations in structural biology has been the development of X-ray free electron lasers (XFELs). The SPring-8 Angstrom Compact free electron LAser (SACLA) in Japan generates the XFEL beam by accelerating electrons to relativistic speeds and directing them through in-vacuum, short-period undulators. Since user operation started in 2012, we have been involved in the development of serial femtosecond crystallography (SFX) measurement systems using XFEL at the SACLA. The SACLA generates X-rays a billion times brighter than SPring-8. The extremely bright XFEL pulses enable data collection with microcrystals (ca. 50–1 μm). Although many molecular analysis techniques exist, SFX is the only technique that can visualize radiation-damage-free structures of biological macromolecules at room temperature in atomic resolution and fast time resolution. Here, we review the achievements of the SACLA-SFX Project in the past 5 years. In particular, we focus on: (1) the measurement system for SFX; (2) experimental phasing by SFX; (3) enzyme chemistry based on damage-free room-temperature structures; and (4) molecular movie taken by time-resolved SFX.