title: Proteomic analysis and identification of Streptococcus pyogenes surface-associated proteins.
abstract: Streptococcus pyogenes is a gram-positive human pathogen that causes a wide spectrum of disease, placing a significant burden on public health. Bacterial surface-associated proteins play crucial roles in host-pathogen interactions and pathogenesis and are important targets for the immune system. The identification of these proteins for vaccine development is an important goal of bacterial proteomics. Here we describe a method of proteolytic digestion of surface-exposed proteins to identify surface antigens of S. pyogenes. Peptides generated by trypsin digestion were analyzed by multidimensional tandem mass spectrometry. This approach allowed the identification of 79 proteins on the bacterial surface, including 14 proteins containing cell wall-anchoring motifs, 12 lipoproteins, 9 secreted proteins, 22 membrane-associated proteins, 1 bacteriophage-associated protein, and 21 proteins commonly identified as cytoplasmic. Thirty-three of these proteins have not been previously identified as cell surface associated in S. pyogenes. Several proteins were expressed in Escherichia coli, and the purified proteins were used to generate specific mouse antisera for use in a whole-cell enzyme-linked immunosorbent assay. The immunoreactivity of specific antisera to some of these antigens confirmed their surface localization. The data reported here will provide guidance in the development of a novel vaccine to prevent infections caused by S. pyogenes.
authors.affiliation: Wyeth Vaccine Research, Pearl River, NY 10965, USA.
authors.forename: Anatoly
authors.initials: A
authors.lastname: Severin
authors.forename: Elliott
authors.initials: E
authors.lastname: Nickbarg
authors.forename: Joseph
authors.initials: J
authors.lastname: Wooters
authors.forename: Shakey A
authors.initials: SA
authors.lastname: Quazi
authors.forename: Yury V
authors.initials: YV
authors.lastname: Matsuka
authors.forename: Ellen
authors.initials: E
authors.lastname: Murphy
authors.forename: Ioannis K
authors.initials: IK
authors.lastname: Moutsatsos
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Zagursky
authors.forename: Stephen B
authors.initials: SB
authors.lastname: Olmsted
mesh_terms: D001426:Bacterial Proteins / Q000032:analysis*; D003593:Cytoplasm / Q000737:chemistry; D004797:Enzyme-Linked Immunosorbent Assay; D008565:Membrane Proteins / Q000032:analysis*; D040901:Proteomics / Q000379:methods*; D013297:Streptococcus pyogenes / Q000737:chemistry*; D014357:Trypsin / Q000494:pharmacology
chemical_list: D001426:Bacterial Proteins; D008565:Membrane Proteins; D014357:Trypsin