Serial femtosecond crystallography at the SACLA: breakthrough to dynamic structural biology
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.
KeywordsDe novo phasing Membrane protein Detergent Bioinorganic chemistry Structure–function relationship Time-resolved analysis
We appreciate all the members of the SACLA-SFX Project, especially Dr. Keitaro Yamashita for the valuable comments on the manuscript. This work was supported by the X-ray Free Electron Laser Priority Strategy Program of the Ministry of Education, Culture, Sports, Science and Technology in Japan and partially by the Strategic Basic Research Programs of the Japan Science and Technology Agency.
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Conflict of interest
Eiichi Mizohata declares that he has no conflict of interest. Takanori Nakane declares that he has no conflict of interest. Yohta Fukuda declares that he has no conflict of interest. Eriko Nango declares that she has no conflict of interest. So Iwata declares that he has no conflict of interest.
This article does not contain any studies with human participants or animals performed by any of the authors.
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