Membrane protein megahertz crystallography at the European XFEL
Membrane protein megahertz crystallography at the European XFEL
复制标题
DOI：
10.1038/s41467-019-12955-3
复制
发表时间：
2019-11-04
影响因子：
16.6
通讯作者：
Zatsepin, Nadia A.
中科院分区：
文献类型：
--
作者：
Gisriel, Chris;Coe, Jesse;Zatsepin, Nadia A.
世界上第一台超导兆赫重复率硬X射线自由电子激光器（XFEL）欧洲XFEL于2017年开始运行，具有独特的脉冲串结构，脉冲之间为886 ns。凭借其快速的脉冲速率，欧洲XFEL可以缓解对XFEL束时间日益增长的需求，特别是对于膜蛋白连续飞秒晶体学（SFX），利用数量级更快的数据收集。在这里，我们报告了第一个膜蛋白兆赫SFX实验，在那里我们确定了一个2.9埃分辨率SFX结构的大膜蛋白复合物，光系统I，一个> 1 MDa复合物含有36个蛋白亚基和381个辅因子。我们解决了兆赫SFX膜蛋白复合物的挑战，包括大量晶体的生长以及影响数据收集和分析的大分子和单位细胞大小。结果表明，兆赫晶体学可能有一个重要的影响，大蛋白质复合物的结构测定与XFEL。
The world's first superconducting megahertz repetition rate hard X-ray free-electron laser (XFEL), the European XFEL, began operation in 2017, featuring a unique pulse train structure with 886 ns between pulses. With its rapid pulse rate, the European XFEL may alleviate some of the increasing demand for XFEL beamtime, particularly for membrane protein serial femtosecond crystallography (SFX), leveraging orders-of-magnitude faster data collection. Here, we report the first membrane protein megahertz SFX experiment, where we determined a 2.9 angstrom-resolution SFX structure of the large membrane protein complex, Photosystem I, a > 1 MDa complex containing 36 protein subunits and 381 cofactors. We address challenges to megahertz SFX for membrane protein complexes, including growth of large quantities of crystals and the large molecular and unit cell size that influence data collection and analysis. The results imply that megahertz crystallography could have an important impact on structure determination of large protein complexes with XFELs.