MRI Consortium: Development of a Damping Wiggler Beamline for X-Ray Footprinting at NSLS II

这项奖励将资助位于布鲁克海文国家实验室的国家同步加速器光源II(NSLS-II)的X射线同步加速器足迹光束线的开发。X射线同步加速器足迹技术(XFP)是一种快速发展的技术，它利用来自同步辐射光源的强烈、电离的白色X射线来产生溶剂衍生的羟基自由基，这些羟基自由基很容易裂解核酸的磷酸二酯骨架，并以共价方式修饰蛋白质中的氨基酸侧链。这种化学的微秒时间尺度使XFP成为探测大分子结构动力学的理想选择，例如多蛋白质复合体的组装、蛋白质折叠过程或配体结合时蛋白质结构的变化。对日益复杂的生理系统进行评估的科学需求已经超过了NSLS现有X28C XFP光束线设施的技术能力，这也将受到计划到2015年用NSLS-II取代NSLS的影响。新的NSLS-II XFP“摇摆器”光束线将取代现有设施，并为大幅增加的信噪比提供更大的白束X射线通量密度，以推动使用现有技术无法实现的新颖和具有挑战性的足迹实验。这些包括水与大分子相互作用的动力学的微秒时间分辨研究，溶液中大分子络合物的表征，膜蛋白质结构分析，以及大分子结构和动力学的活体研究。该计划将增强国际用户社区可用的设施，同时使用先进的NSLS-II技术来推动目前不可行的结构生物学研究。现有的XFP计划服务于40多名资金雄厚的知名研究人员，其中大多数人有许多博士后实习生和研究生，他们可能会在未来的职业生涯中使用这种方法。同步加速器XFP技术的发展与积极的推广和传播计划相结合，已经影响了正在美国各地迅速扩展的非同步加速器足迹和结构质谱学计划，包括该技术在生物技术开发中的商业应用，从而反映了该技术不断扩大的覆盖范围和广泛的用途。NSLS-II XFP光束线设施将是X射线同步加速器足迹的独特国际资源。

This award will fund the development of an X-ray synchrotron footprinting beamline located at the National Synchrotron Light Source-II (NSLS-II) at Brookhaven National Laboratory. X-ray synchrotron footprinting (XFP) is a rapidly growing technique that employs intense, ionizing white X-ray light from a synchrotron source to produce solvent-derived hydroxyl radicals that readily cleave the phosphodiester backbone of nucleic acids and covalently modify amino acid side chains in proteins. The microsecond timescales for this chemistry make XFP ideal for probing macromolecular structural dynamics, such as assembly of multi-protein complexes, protein folding process, or changes in protein structure upon ligand binding. Scientific demand for evaluation of increasingly complex physiological systems has exceeded the technical capabilities of the existing X28C XFP beamline facility at NSLS, which will also be affected by the planned replacement of NSLS with NSLS-II by 2015. The new NSLS-II XFP "wiggler" beamline will replace the existing facility and also provide significantly greater white-beam X-ray flux densities for the substantially increased signal to noise required to drive novel and challenging footprinting experiments unattainable using existing technology. These include microsecond time-resolved studies of the dynamics of water in its interaction with macromolecules, characterization of large megaDalton complexes in solution, membrane protein structural analysis, and in vivo studies of macromolecular structure and dynamics.This program will enhance facilities available to the international user community, while using advanced NSLS-II technologies to drive structural biology studies that are not currently feasible.The existing XFP program serves over forty well-funded established investigators, most of whom have numerous postdoctoral trainees and graduate students who will likely utilize the method in their future careers. Developments in synchrotron XFP technologies coupled to aggressive outreach and dissemination programs have influenced non-synchrotron footprinting and structural mass spectrometry programs that are rapidly expanding across the United States, including commercial application of the technology in biotechnology development, thus reflecting the expanding reach and broad utility of the technique. The NSLS-II XFP beamline facility will represent a unique international resource for x-ray synchrotron footprinting.