title: A gel-free proteomic-based method for the characterization of Bordetella pertussis clinical isolates.
abstract: Bordetella pertussis (Bp) is the etiologic agent of pertussis or whooping cough, a highly contagious respiratory disease occurring primarily in infants and young children. Although vaccine preventable, pertussis cases have increased over the years leading researchers to re-evaluate vaccine control strategies. Since bacterial outer membrane proteins, comprising the surfaceome, often play roles in pathogenesis and antibody-mediated immunity, three recent Bp circulating isolates were examined using proteomics to identify any potential changes in surface protein expression. Fractions enriched for outer membrane proteins were digested with trypsin and the peptides analyzed by nano liquid chromatography-electrospray ionization-mass spectrometry (nLC-ESI-MS), followed by database analysis to elucidate the surfaceomes of our three Bp isolates. Furthermore, a less labor intensive non-gel based antibody affinity capture technology in conjunction with MS was employed to assess each Bp strains' immunogenic outer membrane proteins. This novel technique is generally applicable allowing for the identification of immunogenic surface expressed proteins on pertussis and other pathogenic bacteria.
authors.affiliation: Division of Laboratory Sciences, National Center for Environmental Health, Centers for Disease Control and Prevention, Chamblee, Georgia 30341, USA.
authors.forename: Yulanda M
authors.initials: YM
authors.lastname: Williamson
authors.forename: Hercules
authors.initials: H
authors.lastname: Moura
authors.forename: Kaneatra
authors.initials: K
authors.lastname: Simmons
authors.forename: Jennifer
authors.initials: J
authors.lastname: Whitmon
authors.forename: Nikkol
authors.initials: N
authors.lastname: Melnick
authors.forename: Jon
authors.initials: J
authors.lastname: Rees
authors.forename: Adrian
authors.initials: A
authors.lastname: Woolfitt
authors.forename: David M
authors.initials: DM
authors.lastname: Schieltz
authors.forename: Maria L
authors.initials: ML
authors.lastname: Tondella
authors.forename: Edwin
authors.initials: E
authors.lastname: Ades
authors.forename: Jacquelyn
authors.initials: J
authors.lastname: Sampson
authors.forename: George
authors.initials: G
authors.lastname: Carlone
authors.forename: John R
authors.initials: JR
authors.lastname: Barr
mesh_terms: D000818:Animals; D000907:Antibodies, Bacterial / Q000378:metabolism; D000942:Antigens, Bacterial / Q000302:isolation & purification / Q000378:metabolism; D001426:Bacterial Proteins / Q000032:analysis* / Q000302:isolation & purification; D001886:Bordetella pertussis / Q000737:chemistry* / Q000302:isolation & purification; D002675:Child, Preschool; D002853:Chromatography, Liquid / Q000379:methods; D019295:Computational Biology / Q000379:methods; D005260:Female; D006801:Humans; D007223:Infant; D051379:Mice; D008807:Mice, Inbred BALB C; D011485:Protein Binding; D020543:Proteome / Q000032:analysis* / Q000302:isolation & purification; D040901:Proteomics / Q000379:methods*; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D014917:Whooping Cough / Q000382:microbiology*
chemical_list: D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D020543:Proteome