幽门螺杆菌适应性球形变异相关蛋白的筛选及鉴定  

Screening and identification of related proteins of Helicobacter pylori adaptive coccoid variation

作　　者：曾浩[1] 郭刚[1] 毛旭虎[1] 童文德[1] 邹全明[1] 

机构地区：[1]第三军医大学临床微生物学及免疫学教研室,重庆400038

出　　处：《中国病原生物学杂志》2008年第4期251-254,F0004,共5页Journal of Pathogen Biology

基　　金：国家863计划资助课题(No.2003AA215020);国家"九五"重点科技攻关项目(No.96-901-01-54)

摘　　要：目的对幽门螺杆菌(Hp)在有氧胁迫条件下由螺旋体转变成球形体的适应性调节蛋白进行筛选及鉴定。方法运用双向电泳技术比较Hp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7个与Hp球形变异相关的适应性调节蛋白,即翻译延长因子Tu(EF-Tu)、丙酮酸-黄素蛋白氧化还原酶(POR)、黄素氧化还原蛋白(FldA)、尿素酶G(UreG)、热休克蛋白10(Hsp10)烷基化氢化氧化酶(Ahp)、超氧化物歧化酶(SOD)。结论共筛选出7个与Hp球形变异相关的适应性调节蛋白,对有氧胁迫下Hp球形体的形成机制研究和抗Hp药物靶位和疫苗候选靶位的筛选具有参考价值。Objective Screening and identification of adaptive regulatory proteins of Helicobacter pylori（Hp）during conversion from spiral to coccoid form under oxidative stress.Methods Two-dimensional electrophoresis（2-DE） were employed to compare the differential expression proteins between the spiral form and coccoid form of Hp.The differential proteins were digested in gel by enzyme and the mass of generated peptides were measured by matrix assisted laser desorption 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.Results Seven protein spots associated with the formation of Hp coccoid form were identified in gel as namely,translation elongation factor Tu,Pyruvate ferredoxin oxidoreductase,Flavodoxin,urease G,Heat shock protein 10,Alkyl hydroperoxide reductase,Superoxide dismutase.Conclusion Seven adaptive regulatory proteins associated with Hp coccoid variation were identified.This experiment provides an impetus for further researches of the mechanism of Hp from spiral to coccoid form,of development of new treatment strategy and vaccine design.

关 键 词：幽门螺杆菌 球形体 双向电泳 活性氧代谢 

分 类 号：R378.99[医药卫生—病原生物学]