Bohon Jen
Center for Synchrotron Biosciences, Department of Nutrition, Case Western Reserve University, Cleveland, OH, United States.
Protein Pept Lett. 2019;26(1):55-60. doi: 10.2174/0929866526666181128125125.
First developed in the 1990's at the National Synchrotron Light Source, xray synchrotron footprinting is an ideal technique for the analysis of solution-state structure and dynamics of macromolecules. Hydroxyl radicals generated in aqueous samples by intense x-ray beams serve as fine probes of solvent accessibility, rapidly and irreversibly reacting with solvent exposed residues to provide a "snapshot" of the sample state at the time of exposure. Over the last few decades, improvements in instrumentation to expand the technology have continuously pushed the boundaries of biological systems that can be studied using the technique.
Dedicated synchrotron beamlines provide important resources for examining fundamental biological mechanisms of folding, ligand binding, catalysis, transcription, translation, and macromolecular assembly. The legacy of synchrotron footprinting at NSLS has led to significant improvement in our understanding of many biological systems, from identifying key structural components in enzymes and transporters to in vivo studies of ribosome assembly. This work continues at the XFP (17-BM) beamline at NSLS-II and facilities at ALS, which are currently accepting proposals for use.
X射线同步辐射足迹技术于20世纪90年代在国家同步辐射光源首次开发,是分析大分子溶液状态结构和动力学的理想技术。强X射线束在水性样品中产生的羟基自由基可作为溶剂可及性的精细探针,与暴露于溶剂的残基快速且不可逆地反应,以提供暴露时样品状态的“快照”。在过去几十年中,用于扩展该技术的仪器改进不断推动了可使用该技术研究的生物系统的边界。
专用的同步辐射光束线为研究折叠、配体结合、催化、转录、翻译和大分子组装等基本生物学机制提供了重要资源。NSLS的同步辐射足迹技术遗产使我们对许多生物系统的理解有了显著改善,从识别酶和转运蛋白中的关键结构成分到核糖体组装的体内研究。这项工作在NSLS-II的XFP(17-BM)光束线和ALS的设施中继续进行,目前正在接受使用提案。