title: Proteome analysis of secreted proteins of the gastric pathogen Helicobacter pylori.
abstract: Secreted proteins (the secretome) of the human pathogen Helicobacter pylori may mediate important pathogen-host interactions, but such proteins are technically difficult to analyze. Here, we report on a comprehensive secretome analysis that uses protein-free culture conditions to minimize autolysis, an efficient recovery method for extracellular proteins, and two-dimensional gel electrophoresis followed by peptide mass fingerprinting for protein resolution and identification. Twenty-six of the 33 separated secreted proteins were identified. Among them were six putative oxidoreductases that may be involved in the modification of protein-disulfide bonds, three flagellar proteins, three defined fragments of the vacuolating toxin VacA, the serine protease HtrA, and eight proteins of unknown function. A cleavage site for the amino-terminal passenger domain of VacA between amino acids 991 and 992 was determined by collision-induced dissociation mass spectrometry. Several of the secreted proteins are interesting targets for antimicrobial chemotherapy and vaccine development.
authors.affiliation: Department of Molecular Biology, Max-Planck-Institute for Infection Biology, Berlin, Germany.
authors.forename: Dirk
authors.initials: D
authors.lastname: Bumann
authors.forename: Sevil
authors.initials: S
authors.lastname: Aksu
authors.forename: Meike
authors.initials: M
authors.lastname: Wendland
authors.forename: Katharina
authors.initials: K
authors.lastname: Janek
authors.forename: Uschi
authors.initials: U
authors.lastname: Zimny-Arndt
authors.forename: Nicolas
authors.initials: N
authors.lastname: Sabarth
authors.forename: Thomas F
authors.initials: TF
authors.lastname: Meyer
authors.forename: Peter R
authors.initials: PR
authors.lastname: Jungblut
mesh_terms: D001426:Bacterial Proteins / Q000032:analysis* / Q000378:metabolism; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D016480:Helicobacter pylori / Q000737:chemistry*; D020543:Proteome / Q000032:analysis*; D013270:Stomach / Q000382:microbiology
chemical_list: D001426:Bacterial Proteins; D020543:Proteome