有氧胁迫下幽门螺杆菌球形体形成的比较蛋白质组学研究

Comparative proteomics analysis of Helicobacter pylori from spiral to coccoid form under oxidative stress

目的研究有氧胁迫下幽门螺杆菌(H.pylori)由螺旋体转变成球形体的适应性调节蛋白。方法运用双向电泳技术比较H.pylori螺旋体与球形体的全菌蛋白表达谱,差异蛋白进行胶内酶切,肽混合物使用基质辅助激光解吸/电离飞行时间质谱仪(MALDI-TOF-MS)进行质谱分析,将肽质量指纹谱数据输入互联网上的蛋白质数据库进行检索。结果获得6个与H.pylori球形体形成相关的适应性调节蛋白,即翻译延长因子Tu、丙酮酸-黄素蛋白氧化还原酶、黄素氧化还原蛋白、尿素酶G、烷基化氢化氧化酶、超氧化物歧化酶。结论上述蛋白的鉴定对深入研究有氧胁迫下H.pylori球形体的形成机制,筛选具有潜在价值的抗H.pylori药物靶位和疫苗候选表位具有参考价值。

[Objective] To study protein variation of H.pylori during conversion from spiral to coecoid form under oxidative stress. [Methods] Two-dimensional electrophoresis （2-DE） were used to compare the differential expression proteins between the spiral form and coecoid form of H.pylori. The differential proteins were digested in gel by enzyme and the mass of generated peptides were measured by matrix assisted laser desorpfion ionization time of flight mass spectrometry （MALDI-TOF-MS）. The data obtained from peptide mass finger-printing （PMF） were searched using the internet available database. [Result] Six spots proteins associated with the formation of H.pylori coccoid form were identified in gel as namely, translation elongation factor Tu, pyruvate ferredoxin oxidoreductase, flavodoxin, urease G, alkyl hydroperoxide reductase, superoxide dismutase. [Conclusion] This experiment provides an impetus for further researches of the mechanism of H.pylori from spiral to coccoid form under oxidative stress and a development of new treatment strategy and vaccine bolting.