Serial femtosecond crystallography: the first five years.
Serial femtosecond crystallography: the first five years.
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DOI：
10.1107/s205225251402702x
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发表时间：
2015-03-01
期刊：
影响因子：
3.9
通讯作者：
Schlichting I
中科院分区：
文献类型：
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作者：
Schlichting I
硬X射线自由电子激光的出现为大分子晶体学揭开了新的篇章。本文介绍了连续飞秒晶体学的最新研究成果、进展和展望。使用同步辐射源的蛋白质晶体学对生物学产生了巨大的影响，产生了数千种蛋白质的结构，并详细了解了它们的机制。然而，该技术受限于对宏观晶体的要求，这可能很难获得，以及在衍射实验中造成的通常严重的辐射损伤，特别是当使用微小晶体时。为了减缓辐射损伤，数据收集通常在低温下进行。随着自由电子激光器（FEL）能够提供极强的飞秒X射线脉冲的出现，这种情况似乎得到了补救，允许使用宏观或微观晶体来确定未受损的大分子的结构。后者暴露于自由电子激光束在随机取向和它们的衍射数据收集在低温或室温下在一个串行的方式，因为每个晶体被破坏后，一个单一的曝光。晶体生长和交付，衍射数据分析，包括从头定相，所需的新方法进行审查。SFX的机遇和挑战，包括应用程序，如时间分辨测量和分析的辐射损伤倾向的系统。
The advent of hard X-ray free-electron lasers has opened a new chapter in macromolecular crystallography. Recent results, developments and prospects of serial femtosecond crystallography are described. Protein crystallography using synchrotron radiation sources has had a tremendous impact on biology, having yielded the structures of thousands of proteins and given detailed insight into their mechanisms. However, the technique is limited by the requirement for macroscopic crystals, which can be difficult to obtain, as well as by the often severe radiation damage caused in diffraction experiments, in particular when using tiny crystals. To slow radiation damage, data collection is typically performed at cryogenic temperatures. With the advent of free-electron lasers (FELs) capable of delivering extremely intense femtosecond X-ray pulses, this situation appears to be remedied, allowing the structure determination of undamaged macromolecules using either macroscopic or microscopic crystals. The latter are exposed to the FEL beam in random orientations and their diffraction data are collected at cryogenic or room temperature in a serial fashion, since each crystal is destroyed upon a single exposure. The new approaches required for crystal growth and delivery, and for diffraction data analysis, including de novo phasing, are reviewed. The opportunities and challenges of SFX are described, including applications such as time-resolved measurements and the analysis of radiation damage-prone systems.