Introduction:
Synchrotron X-ray mediated hydroxyl radical footprinting provides information about the structure and dynamics of biological macromolecules in solution. In this method an intense X-ray from a bending magnet source is used to generate a burst of hydroxyl radicals (in millisecond timescale), which interact with the solvent accessible sites of a nucleic acid or a protein. The locations and the extent of strand cleavages in the nucleic acid and the numerous sites of covalent modifications in the amino acid side chains are determined by gel electrophoresis (Figure 1) and mass spectrometry (Figure 2) respectively. The extent of the hydroxyl radical reaction is correlated directly to the solvent accessibilities of the regions. Experiments are carried out with micro or sub-micromolar concentrations of biological macromolecules at physiological conditions. For past several years synchrotron footprinting method has been carried out to investigate RNA folding, protein-protein and protein-nucleic acid interactions at a resolution of a single nucleoside or amino-acid level. Recent technical and analytical advancements have opened a new approach to examine the structure of large complexes, membrane proteins and bimolecular interactions inside the living cell by synchrotron footprinting methodology.
Research Programs
The research team of CSB-X28C, designs, tests, and implements the progress made in the hardware and the software technologies associated with synchrotron footprinting for biomedical applications. Recent research projects include probing structure and function of mega Dalton bimolecular complexes, membrane proteins and in vivo studies using steady-state and time-resolved synchrotron footprinting. Moreover, CSB-X28C provides a state-of-the-art laboratory for synchrotron footprinting research with scientific, experimental and technical assistance to users from all over the world. CSB research facility carries out core, collaborative and service research projects. The core research program strives to continually re-discover new experimental and analytical technologies in synchrotron footprinting so as to emphasize its uniqueness and prominence in the field of Structural Biology. For past several years, CSB-X28C has been doing collaborative work with an array of users around the globe. The collaborative research programs are specifically developed to drive the core research programs. The service research programs are designed for a user-friendly, rapid and cost -free access to the facility.
User Support
The CSB-X28C provides user support for research projects conducted by academic, government and industrial institutions. CSB-X28C also provides comprehensive beamline support to the general users of NSLS.
PUBLICATIONS
