Ghosh Swagatha, Zorić Doris, Dahl Peter, Bjelčić Monika, Johannesson Jonatan, Sandelin Emil, Borjesson Per, Björling Alexander, Banacore Analia, Edlund Petra, Aurelius Oskar, Milas Mirko, Nan Jie, Shilova Anastasya, Gonzalez Ana, Mueller Uwe, Brändén Gisela, Neutze Richard
Department of Chemistry and Molecular Biology, University of Gothenburg, Medicinaregatan 9C, 40530 Gothenburg, Sweden.
MAX IV Laboratory, Lund University, Fotongatan 2, 224 84 Lund, Sweden.
J Appl Crystallogr. 2023 Mar 9;56(Pt 2):449-460. doi: 10.1107/S1600576723001036. eCollection 2023 Apr 1.
Serial femtosecond crystallography was initially developed for room-temperature X-ray diffraction studies of macromolecules at X-ray free electron lasers. When combined with tools that initiate biological reactions within microcrystals, time-resolved serial crystallography allows the study of structural changes that occur during an enzyme catalytic reaction. Serial synchrotron X-ray crystallography (SSX), which extends serial crystallography methods to synchrotron radiation sources, is expanding the scientific community using serial diffraction methods. This report presents a simple flow cell that can be used to deliver microcrystals across an X-ray beam during SSX studies. This device consists of an X-ray transparent glass capillary mounted on a goniometer-compatible 3D-printed support and is connected to a syringe pump via light-weight tubing. This flow cell is easily mounted and aligned, and it is disposable so can be rapidly replaced when blocked. This system was demonstrated by collecting SSX data at MAX IV Laboratory from microcrystals of the integral membrane protein cytochrome oxidase from , from which an X-ray structure was determined to 2.12 Å resolution. This simple SSX platform may help to lower entry barriers for non-expert users of SSX.
串行飞秒晶体学最初是为在X射线自由电子激光下对大分子进行室温X射线衍射研究而开发的。当与能引发微晶内生物反应的工具相结合时,时间分辨串行晶体学能够研究酶催化反应过程中发生的结构变化。串行同步辐射X射线晶体学(SSX)将串行晶体学方法扩展到同步辐射源,正在扩大使用串行衍射方法的科学界。本报告介绍了一种简单的流动池,可用于在SSX研究期间将微晶输送穿过X射线束。该装置由安装在与测角仪兼容的3D打印支架上的X射线透明玻璃毛细管组成,并通过轻质 tubing 连接到注射泵。这种流动池易于安装和对准,并且是一次性的,因此在堵塞时可以快速更换。通过在MAX IV实验室从嗜热栖热菌的整合膜蛋白细胞色素c氧化酶的微晶中收集SSX数据证明了该系统,从中确定了分辨率为2.12 Å的X射线结构。这个简单的SSX平台可能有助于降低SSX非专业用户的入门门槛。