BioCARS, Center for Advanced Radiation Sources, The University of Chicago, 9700 South Cass Ave, Bld 434B, Lemont, IL 60439, USA.
Department of Chemistry, Korea Advanced Institute of Science and Technology, E6-6 #513, 291 Daehak-ro, Daejeon, Yuseong-gu 34141, Republic of Korea.
J Synchrotron Radiat. 2023 Mar 1;30(Pt 2):490-499. doi: 10.1107/S1600577522012127. Epub 2023 Feb 1.
A fundamental problem in biological sciences is understanding how macromolecular machines work and how the structural changes of a molecule are connected to its function. Time-resolved techniques are vital in this regard and essential for understanding the structural dynamics of biomolecules. Time-resolved small- and wide-angle X-ray solution scattering has the capability to provide a multitude of information about the kinetics and global structural changes of molecules under their physiological conditions. However, standard protocols for such time-resolved measurements often require significant amounts of sample, which frequently render time-resolved measurements impossible. A cytometry-type sheath co-flow cell, developed at the BioCARS 14-ID beamline at the Advanced Photon Source, USA, allows time-resolved pump-probe X-ray solution scattering measurements to be conducted with sample consumption reduced by more than ten times compared with standard sample cells and protocols. The comparative capabilities of the standard and co-flow experimental setups were demonstrated by studying time-resolved signals in photoactive yellow protein.
生物科学中的一个基本问题是理解大分子机器如何工作以及分子的结构变化如何与其功能相关联。在这方面,时间分辨技术至关重要,是理解生物分子结构动力学的关键。时间分辨小角和宽角 X 射线溶液散射技术有能力提供关于分子在生理条件下的动力学和整体结构变化的大量信息。然而,此类时间分辨测量的标准方案通常需要大量的样品,这经常使得时间分辨测量变得不可能。在美国先进光源的 BioCARS 14-ID 光束线上开发的一种细胞仪型鞘流池,使时间分辨泵浦探测 X 射线溶液散射测量能够以比标准样品池和方案减少超过十倍的样品消耗来进行。通过研究光活性黄色蛋白中的时间分辨信号,证明了标准和共流实验装置的比较能力。