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New high-throughput endstation to accelerate the experimental optimization pipeline for synchrotron X-ray footprinting
Journal of Synchrotron Radiation ( IF 2.5 ) Pub Date : 2021-07-20 , DOI: 10.1107/s1600577521005026
Rohit Jain 1 , Donald Abel 1 , Maksim Rakitin 2 , Michael Sullivan 1 , David T Lodowski 3 , Mark R Chance 1 , Erik R Farquhar 1
Affiliation  

Synchrotron X-ray footprinting (XF) is a growing structural biology technique that leverages radiation-induced chemical modifications via X-ray radiolysis of water to produce hydroxyl radicals that probe changes in macromolecular structure and dynamics in solution states of interest. The X-ray Footprinting of Biological Materials (XFP) beamline at the National Synchrotron Light Source II provides the structural biology community with access to instrumentation and expert support in the XF method, and is also a platform for development of new technological capabilities in this field. The design and implementation of a new high-throughput endstation device based around use of a 96-well PCR plate form factor and supporting diagnostic instrumentation for synchrotron XF is described. This development enables a pipeline for rapid comprehensive screening of the influence of sample chemistry on hydroxyl radical dose using a convenient fluorescent assay, illustrated here with a study of 26 organic compounds. The new high-throughput endstation device and sample evaluation pipeline now available at the XFP beamline provide the worldwide structural biology community with a robust resource for carrying out well optimized synchrotron XF studies of challenging biological systems with complex sample compositions.

中文翻译:

新型高通量终端站可加速同步加速器 X 射线足迹实验优化流程

同步加速器 X 射线足迹 (XF) 是一种不断发展的结构生物学技术,它通过水的 X 射线辐射分解,利用辐射诱导的化学修饰产生羟基自由基,从而探测感兴趣的溶液状态中大分子结构和动力学的变化。国家同步加速器光源 II 的生物材料 X 射线足迹 (XFP) 光束线为结构生物学界提供了 XF 方法中的仪器和专家支持,也是该领域新技术能力开发的平台。描述了基于 96 孔 PCR 板形状因素和同步加速器 XF 支持诊断仪器的新型高通量终端站设备的设计和实现。这一发展使得使用方便的荧光测定快速全面筛选样品化学对羟基自由基剂量的影响成为可能,此处通过对 26 种有机化合物的研究进行了说明。XFP 光束线现已提供新型高通量终端站设备和样品评估管道,为全球结构生物学界提供了强大的资源,用于对具有复杂样品成分的挑战性生物系统进行优化的同步加速器 XF 研究。
更新日期:2021-09-03
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