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Mutation of cytotoxin-associated gene A affects expressions of antioxidant proteins of helicobacter pylori

World Journal of Gastroenterology 2009年第5期15卷 599-606页

作者： Zhi-Gang Huang Guang-Cai Duan Qing-Tang Fan Wei-Dong Zhang Chun-Hua Song Xue-Yong Huang Rong-Guang Zhang Department of Epidemiology College of Public HealthZhengzhou University Zhengzhou 450001 Henan ProvinceChina Department of Epidemiology School of Public Health Guangdong Medical College Dongguan 523808Guangdong Province China

AIM: To determine if disruption of the cagA gene of helicobacter pylori (H pylori) has an effect on the expression of other proteins at proteome level. METHODS: Construction of a cagA knock out mutant Hp27 _△cagA (cagA -) via homologous recombination with the wild-type strain Hp27 (cagA+) as a recipient was performed. The method of sonication-urea-CHAPS-DTT was employed to extract bacterial proteins from both strains. Soluble proteins were analyzed by two-dimensional electrophoresis (2-DE). Images of 2-DE gels were digitalized and analyzed. Only spots that had a statistical signif icance in differential expression were selected and analyzed by matrix-assisted laser desorption/ionizationtime of flight mass spectrometry (MALDI-TOF-MS). Biological information was used to search protein database and identify the biological function of proteins. RESULTS: The proteome expressions between wild-type strain and isogenic mutant with the cagA gene knocked-out were compared. Five protein spots with high abundance in bacteria proteins of wild-type strains, down-regulated or absently expressed in bacteria proteins of mutants, were identified and analyzed. From a quantitative point of view, the identified proteins are related to the cagA gene and important antioxidant proteins of H pylori , including alkyl hydroperoxide reductase (Ahp), superoxide dismutase (SOD) and modulator of drug activity (Mda66), respectively, suggesting that cagA is important to maintain the normal activity of antioxidative stress and ensure H pylori persistent colonization in the host. CONCLUSION: cagA gene is relevant to the expressions of antioxidant proteins of H pylori, which may be a novel mechanism involved in H pylori cagA pathogenesis.

关键词： helicobacter pylorr, cytotoxin-associatedgene a Knock-out Antioxidant protein Two-dimensional electrophoresis

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