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

Case Western Reserve University, Cleveland OH

Investigators

Abstract

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.