Proteomic analysis of proteins expressed by Helicobacter pylori under oxidative stress.
Helicobacter pylori is a spiral, slow growing gram-negative microaerophilic bacterium. It has been shown to be the etiological agent of gastroduodenal diseases, such as chronic gastritis, gastric and duodenal ulcers, and gastric cancer. To address the influence of oxidative stress and its underlying mechanisms, we have compared proliferation, urease activity and protein expression profile of H. pylori incubated under normal microaerophilic (5% O2) and aerobic stress (20% O2) conditions. Oxidative-stress cells displayed coccoid morphology and time-dependent decrease in proliferation. The urease activity was completely abrogated after 32 h. We have further compared the protein expression profiles of H. pylori under normal growing and oxidative-stress conditions by a global proteomic analysis, which includes high-resolution 2-DE followed by MALDI-TOF-MS and bioinformatic databases search/peptide-mass comparison. The results revealed that more than ten proteins were differentially expressed under oxidative stress. Most notably, the protein expression levels of urease accessory protein E (UreE, an essential metallochaperone for urease activity) and alkylhydroperoxide reductase (AhpC) with antioxidant potential are greatly decreased under stress conditions. Measurements of messenger RNA transcription level by performing RT-PCR on total mRNA also confirmed that gene expressions for these two proteins are consistently repressed under oxygen tension. These changes form a firm basis to account for the loss of urease activity and anti-oxidative ability of H. pylori after long-term exposure to reactive oxygen. Conceivably, UreE and AhpC may thus be listed as potential targets for the development of therapeutic drugs against H. pylori.
Chuang
Ming-Hong
MH
Institute of Biochemical Sciences, National Taiwan University, Taipei, Taiwan.
Wu
Ming-Shiang
MS
Lin
Jou-Town
JT
Chiou
Shyh-Horng
SH
D000818:Animals; D049109:Cell Proliferation; D015180:Electrophoresis, Gel, Two-Dimensional; D016480:Helicobacter pylori / Q000166:cytology / Q000378:metabolism*; D018384:Oxidative Stress; D011506:Proteins / Q000378:metabolism*; D020543:Proteome; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014510:Urease / Q000378:metabolism
D011506:Proteins; D020543:Proteome; D014510:Urease