title: Structural proteomics of macromolecular assemblies using oxidative footprinting and mass spectrometry.
abstract: Understanding the composition, structure and dynamics of macromolecules and their assemblies is at the forefront of biological science today. Hydroxyl-radical-mediated protein footprinting using mass spectrometry can define macromolecular structure, macromolecular assembly and conformational changes of macromolecules in solution based on measurements of reactivity of amino acid side-chain groups with covalent-modification reagents. Subsequent to oxidation by reactive oxygen species, proteins are digested by specific proteases to generate peptides for analysis by mass spectrometry. Accurate measurements of side-chain reactivity are achieved using quantitative liquid-chromatography-coupled mass spectrometry, whereas the side-chain sites within the macromolecular probes are identified using tandem mass spectrometry. In addition, the use of footprinting data in conjunction with computational modeling approaches is a powerful new method for testing and refining structural models of macromolecules and their complexes.
authors.affiliation: Case Center for Proteomics and Mass Spectrometry, 930 BRB, Case Western Reserve University, 10900 Euclid Avenue, Cleveland, OH 44106, USA.
authors.forename: Jing-Qu
authors.initials: JQ
authors.lastname: Guan
authors.forename: Mark R
authors.initials: MR
authors.lastname: Chance
mesh_terms: D002853:Chromatography, Liquid; D019295:Computational Biology; D017665:Hydroxyl Radical; D046911:Macromolecular Substances; D013058:Mass Spectrometry; D008961:Models, Structural; D010084:Oxidation-Reduction; D011487:Protein Conformation; D023302:Protein Footprinting; D011506:Proteins / Q000378:metabolism; D040901:Proteomics; D017382:Reactive Oxygen Species; D013329:Structure-Activity Relationship
chemical_list: D046911:Macromolecular Substances; D011506:Proteins; D017382:Reactive Oxygen Species; D017665:Hydroxyl Radical