title: Complete protein characterization using top-down mass spectrometry and ultraviolet photodissociation.
abstract: The top-down approach to proteomics offers compelling advantages due to the potential to provide complete characterization of protein sequence and post-translational modifications. Here we describe the implementation of 193 nm ultraviolet photodissociation (UVPD) in an Orbitrap mass spectrometer for characterization of intact proteins. Near-complete fragmentation of proteins up to 29 kDa is achieved with UVPD including the unambiguous localization of a single residue mutation and several protein modifications on Pin1 (Q13526), a protein implicated in the development of Alzheimer's disease and in cancer pathogenesis. The 5 ns, high-energy activation afforded by UVPD exhibits far less precursor ion-charge state dependence than conventional collision- and electron-based dissociation methods.
authors.affiliation: Department of Chemistry and Biochemistry, The University of Texas at Austin, 1 University Station A5300, Austin, Texas 78712, USA.
authors.forename: Jared B
authors.initials: JB
authors.lastname: Shaw
authors.forename: Wenzong
authors.initials: W
authors.lastname: Li
authors.forename: Dustin D
authors.initials: DD
authors.lastname: Holden
authors.forename: Yan
authors.initials: Y
authors.lastname: Zhang
authors.forename: Jens
authors.initials: J
authors.lastname: Griep-Raming
authors.forename: Ryan T
authors.initials: RT
authors.lastname: Fellers
authors.forename: Bryan P
authors.initials: BP
authors.lastname: Early
authors.forename: Paul M
authors.initials: PM
authors.lastname: Thomas
authors.forename: Neil L
authors.initials: NL
authors.lastname: Kelleher
authors.forename: Jennifer S
authors.initials: JS
authors.lastname: Brodbelt
mesh_terms: D006801:Humans; D013058:Mass Spectrometry; D008958:Models, Molecular; D000072340:NIMA-Interacting Peptidylprolyl Isomerase; D019696:Peptidylprolyl Isomerase / Q000032:analysis* / Q000235:genetics; D055668:Photochemical Processes; D040901:Proteomics; D014466:Ultraviolet Rays
chemical_list: D000072340:NIMA-Interacting Peptidylprolyl Isomerase; C000606846:PIN1 protein, human; D019696:Peptidylprolyl Isomerase