Cohen Aina E, Doukov Tzanko, Soltis Michael S
Stanford Synchrotron Radiation Lightsource, SLAC National Accelerator Laboratory, Stanford University, Menlo Park, CA 94025, USA.
Protein Pept Lett. 2016;23(3):283-90. doi: 10.2174/0929866523666160107115015.
This review describes the use of single crystal UV-Visible Absorption micro-Spectrophotometry (UV-Vis AS) to enhance the design and execution of X-ray crystallography experiments for structural investigations of reaction intermediates of redox active and photosensitive proteins. Considerations for UV-Vis AS measurements at the synchrotron and associated instrumentation are described. UV-Vis AS is useful to verify the intermediate state of an enzyme and to monitor the progression of reactions within crystals. Radiation induced redox changes within protein crystals may be monitored to devise effective diffraction data collection strategies. An overview of the specific effects of radiation damage on macromolecular crystals is presented along with data collection strategies that minimize these effects by combining data from multiple crystals used at the synchrotron and with the X-ray free electron laser.
本综述描述了如何使用单晶紫外可见吸收显微分光光度法（UV-Vis AS）来改进X射线晶体学实验的设计与实施，以对氧化还原活性和光敏蛋白的反应中间体进行结构研究。文中介绍了在同步加速器上进行UV-Vis AS测量的注意事项及相关仪器设备。UV-Vis AS有助于验证酶的中间状态，并监测晶体内反应的进程。可以监测蛋白质晶体中辐射诱导的氧化还原变化，以制定有效的衍射数据收集策略。本文概述了辐射损伤对大分子晶体的具体影响，并介绍了通过合并在同步加速器和X射线自由电子激光上使用的多个晶体的数据来最小化这些影响的数据收集策略。
