Shih O, Liao K-F, Yeh Y-Q, Su C-J, Wang C-A, Chang J-W, Wu W-R, Liang C-C, Lin C-Y, Lee T-H, Chang C-H, Chiang L-C, Chang C-F, Liu D-G, Lee M-H, Liu C-Y, Hsu T-W, Mansel B, Ho M-C, Shu C-Y, Lee F, Yen E, Lin T-C, Jeng U
National Synchrotron Radiation Research Center, Hsinchu Science Park, Hsinchu 30076, Taiwan.
Institute of Biochemical Sciences and Institute of Biological Chemistry, Academia Sinica, Nankang, Taipei 11529, Taiwan.
J Appl Crystallogr. 2022 Mar 18;55(Pt 2):340-352. doi: 10.1107/S1600576722001923. eCollection 2022 Apr 1.
Recent developments in the instrumentation and data analysis of synchrotron small-angle X-ray scattering (SAXS) on biomolecules in solution have made biological SAXS (BioSAXS) a mature and popular tool in structural biology. This article reports on an advanced endstation developed at beamline 13A of the 3.0 GeV Taiwan Photon Source for biological small- and wide-angle X-ray scattering (SAXS-WAXS or SWAXS). The endstation features an in-vacuum SWAXS detection system comprising two mobile area detectors (Eiger X 9M/1M) and an online size-exclusion chromatography system incorporating several optical probes including a UV-Vis absorption spectrometer and refractometer. The instrumentation and automation allow simultaneous SAXS-WAXS data collection and data reduction for high-throughput biomolecular conformation and composition determinations. The performance of the endstation is illustrated with the SWAXS data collected for several model proteins in solution, covering a scattering vector magnitude across three orders of magnitude. The crystal-model fittings to the data in the range ∼0.005-2.0 Å indicate high similarity of the solution structures of the proteins to their crystalline forms, except for some subtle hydration-dependent local details. These results open up new horizons of SWAXS in studying correlated local and global structures of biomolecules in solution.
同步加速器小角X射线散射（SAXS）对溶液中生物分子的仪器和数据分析方面的最新进展，使生物SAXS（BioSAXS）成为结构生物学中一种成熟且流行的工具。本文报道了在3.0 GeV台湾光子源的13A光束线开发的用于生物小角和广角X射线散射（SAXS-WAXS或SWAXS）的先进终端站。该终端站的特点是有一个真空内的SWAXS检测系统，包括两个移动面探测器（Eiger X 9M/1M）和一个在线尺寸排阻色谱系统，该系统包含多个光学探头，包括紫外可见吸收光谱仪和折射仪。该仪器和自动化功能允许同时进行SAXS-WAXS数据采集和数据处理，以进行高通量生物分子构象和组成测定。通过为溶液中的几种模型蛋白收集的SWAXS数据说明了该终端站的性能，这些数据覆盖了三个数量级的散射矢量大小。在约0.005-2.0 Å范围内对数据进行的晶体模型拟合表明，除了一些与水合作用相关的细微局部细节外，这些蛋白的溶液结构与其晶体形式具有高度相似性。这些结果为SWAXS研究溶液中生物分子的相关局部和整体结构开辟了新的视野。
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