Proteome analysis of secreted proteins of the gastric pathogen Helicobacter pylori.
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.
Bumann
Dirk
D
Department of Molecular Biology, Max-Planck-Institute for Infection Biology, Berlin, Germany.
Aksu
Sevil
S
Wendland
Meike
M
Janek
Katharina
K
Zimny-Arndt
Uschi
U
Sabarth
Nicolas
N
Meyer
Thomas F
TF
Jungblut
Peter R
PR
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
D001426:Bacterial Proteins; D020543:Proteome