title: Protein Structural Analysis via Mass Spectrometry-Based Proteomics.
abstract: Modern mass spectrometry (MS) technologies have provided a versatile platform that can be combined with a large number of techniques to analyze protein structure and dynamics. These techniques include the three detailed in this chapter: (1) hydrogen/deuterium exchange (HDX), (2) limited proteolysis, and (3) chemical crosslinking (CX). HDX relies on the change in mass of a protein upon its dilution into deuterated buffer, which results in varied deuterium content within its backbone amides. Structural information on surface exposed, flexible or disordered linker regions of proteins can be achieved through limited proteolysis, using a variety of proteases and only small extents of digestion. CX refers to the covalent coupling of distinct chemical species and has been used to analyze the structure, function and interactions of proteins by identifying crosslinking sites that are formed by small multi-functional reagents, termed crosslinkers. Each of these MS applications is capable of revealing structural information for proteins when used either with or without other typical high resolution techniques, including NMR and X-ray crystallography.
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Kansas Medical Center, Kansas City, KS, USA. aartigues@kumc.edu.
authors.forename: Antonio
authors.initials: A
authors.lastname: Artigues
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Kansas Medical Center, Kansas City, KS, USA.
authors.forename: Owen W
authors.initials: OW
authors.lastname: Nadeau
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Kansas Medical Center, Kansas City, KS, USA.
authors.forename: Mary Ashley
authors.initials: MA
authors.lastname: Rimmer
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Kansas Medical Center, Kansas City, KS, USA.
authors.forename: Maria T
authors.initials: MT
authors.lastname: Villar
authors.affiliation: Department of Bioinformatics and Genomics, University of North Carolina at Charlotte, Charlotte, NC, USA.
authors.forename: Xiuxia
authors.initials: X
authors.lastname: Du
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Kansas Medical Center, Kansas City, KS, USA.
authors.forename: Aron W
authors.initials: AW
authors.lastname: Fenton
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Kansas Medical Center, Kansas City, KS, USA.
authors.forename: Gerald M
authors.initials: GM
authors.lastname: Carlson
mesh_terms: D000465:Algorithms; D000818:Animals; D019295:Computational Biology / Q000379:methods*; D003432:Cross-Linking Reagents / Q000737:chemistry; D057225:Data Mining / Q000379:methods*; D030562:Databases, Protein; D041961:Deuterium Exchange Measurement; D057166:High-Throughput Screening Assays; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods*; D011487:Protein Conformation; D011506:Proteins / Q000032:analysis*; D059748:Proteolysis; D020543:Proteome; D040901:Proteomics / Q000379:methods*; D015203:Reproducibility of Results; D012984:Software; D057188:Workflow
chemical_list: D003432:Cross-Linking Reagents; D011506:Proteins; D020543:Proteome
keywords: Chemical Crosslinking (CX); Hydrogen/Deuterium Exchange (HDX); Limited proteolysis; Protein structural analysis