title: Recent Advances and Applications in Synchrotron X-Ray Protein Footprinting for Protein Structure and Dynamics Elucidation.
abstract: Synchrotron X-ray Footprinting is a powerful in situ hydroxyl radical labeling method for analysis of protein structure, interactions, folding and conformation change in solution. In this method, water is ionized by high flux density broad band synchrotron X-rays to produce a steady-state concentration of hydroxyl radicals, which then react with solvent accessible side-chains. The resulting stable modification products are analyzed by liquid chromatography coupled to mass spectrometry. A comparative reactivity rate between known and unknown states of a protein provides local as well as global information on structural changes, which is then used to develop structural models for protein function and dynamics. In this review we describe the XF-MS method, its unique capabilities and its recent technical advances at the Advanced Light Source. We provide a comparison of other hydroxyl radical and mass spectrometry based methods with XFMS. We also discuss some of the latest developments in its usage for studying bound water, transmembrane proteins and photosynthetic protein components, and the synergy of the method with other synchrotron based structural biology methods.
authors.forename: Sayan
authors.initials: S
authors.lastname: Gupta
authors.forename: Jun
authors.initials: J
authors.lastname: Feng
authors.forename: Mark
authors.initials: M
authors.lastname: Chance
authors.affiliation: Molecular Biophysics and Integrated Bioimaging, Experimental Systems, Advanced Light Source, Lawrence Berkeley National Laboratory, Berkeley, CA. CYRalston@ibi.gov.
authors.forename: Corie
authors.initials: C
authors.lastname: Ralston
mesh_terms: D017665:Hydroxyl Radical; D013058:Mass Spectrometry / Q000295:instrumentation; D008958:Models, Molecular; D011487:Protein Conformation; D023302:Protein Footprinting / Q000295:instrumentation*; D011506:Proteins / Q000737:chemistry*; D017356:Synchrotrons; D014965:X-Rays
chemical_list: D011506:Proteins; D017665:Hydroxyl Radical