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

该奖项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法（XFP）是一种快速发展的技术，其采用来自同步加速器源的强电离白色X射线光来产生溶剂衍生的羟基自由基，所述羟基自由基容易地切割核酸的磷酸二酯主链并共价修饰蛋白质中的氨基酸侧链。这种化学反应的微秒时间尺度使XFP成为探测大分子结构动力学的理想选择，例如多蛋白质复合物的组装，蛋白质折叠过程或配体结合后蛋白质结构的变化。评估日益复杂的生理系统的科学需求已经超过了NSLS现有X28 C XFP光束线设施的技术能力，这也将受到计划在2015年前用NSLS-II取代NSLS的影响。新的NSLS-II XFP“摇摆器”光束线将取代现有的设施，并提供显着更大的白束X射线通量密度，以大幅增加驱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所需的信噪比。其中包括水与大分子相互作用的动力学的微秒时间分辨研究，溶液中大分子复合物的表征，膜蛋白结构分析，以及大分子结构和动力学的体内研究。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计划为40多名资金充足的既定研究人员提供服务，他们中的大多数都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技术的发展加上积极的推广和传播计划，影响了在美国迅速扩大的非同步加速器足迹和结构质谱计划，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范围和广泛的实用性。NSLS-II XFP光束线设施将成为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.