title: Proteomics analyses of the opportunistic pathogen Burkholderia vietnamiensis using protein fractionations and mass spectrometry.
abstract: The main objectives of this work were to obtain a more extensive coverage of the Burkholderia vietnamiensis proteome than previously reported and to identify virulence factors using tandem mass spectrometry. The proteome of B. vietnamiensis was precipitated into four fractions to as extracellular, intracellular, cell surface and cell wall proteins. Two different approaches were used to analyze the proteins. The first was a gel-based method where 1D SDS-PAGE was used for separation of the proteins prior to reverse phase liquid chromatography tandem mass spectrometry (LC-MS/MS). The second method used MudPIT analysis (Multi dimensional Protein Identification Technique), where proteins are digested and separated using cation exchange and reversed phase separations before the MS/MS analysis (LC/LC-MS/MS). Overall, gel-based LC-MS/MS analysis resulted in more protein identifications than the MudPIT analysis. Combination of the results lead to identification of more than 1200 proteins, approximately 16% of the proteins coded from the annotated genome of Burkholderia species. Several virulence factors were detected including flagellin, porin, peroxiredoxin and zinc proteases.
authors.affiliation: Department of Chemistry and Biochemistry, University of Arizona, 1306 E University Bolevard, Tucson, AZ 85721, USA.
authors.forename: Samanthi
authors.initials: S
authors.lastname: Wickramasekara
authors.forename: Julie
authors.initials: J
authors.lastname: Neilson
authors.forename: Naren
authors.initials: N
authors.lastname: Patel
authors.forename: Linda
authors.initials: L
authors.lastname: Breci
authors.forename: Amy
authors.initials: A
authors.lastname: Hilderbrand
authors.forename: Raina M
authors.initials: RM
authors.lastname: Maier
authors.forename: Vicki
authors.initials: V
authors.lastname: Wysocki
mesh_terms: D001426:Bacterial Proteins / Q000032:analysis* / Q000378:metabolism*; D019117:Burkholderia / Q000737:chemistry* / Q000378:metabolism*; D002853:Chromatography, Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D020543:Proteome / Q000032:analysis*; D040901:Proteomics / Q000379:methods*; D053719:Tandem Mass Spectrometry
chemical_list: D001426:Bacterial Proteins; D020543:Proteome