幽门螺杆菌球形休眠体形成的比较蛋白质组学研究

Comparative proteomics analysis of Helicobacter pylori from spiral to dormant coccoid form

目的 研究幽门螺杆菌由螺杆状转变成球形休眠体的相关蛋白。方法 运用双向电泳技术比较螺杆状幽门螺杆菌及其球形休眠体全菌蛋白表达谱 ,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 9个与幽门螺杆菌球形休眠体形成相关蛋白 ,即相对分子质量 (Mr)为 2 6× 10 3的过氧化物酶、硫氧还蛋白、烯醇酶、黄素氧化还原蛋白、单链DNA结合蛋白、翻译起始因子 1、延长因子P、中性粒细胞激活蛋白和Mr 为 10× 10 3的热休克蛋白。结论 上述蛋白的鉴定为深入研究幽门螺杆菌球形休眠体形成机制 ,筛选具有潜在价值的抗幽门螺杆菌药物靶位和疫苗候选靶位具有参考价值。

Objective To study protein variation of Helicobacter pylori during conversion from spiral to dormant coccoid form. Methods Two-dimensional electrophoresis (2-DE) were employed to compare the differential expression proteins between the spiral form and dormant coccoid 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 desorption ionization time of flight mass spectrometry (MALDI-TOF-MS). The data obtained from peptide mass fingerprinting (PMF) were searched using the internet available database. Results Nine spots of proteins associated with the formation of domant coccoid form were identified in gel as namely, probable peroxiredoxin ( M _r 26×10 3 antigen), thioredoxin, enolase, flavodoxin, ss-DNA binding protein 12 RNP2 precursor, translation initiation factor IF-1, elongation factor P, neutrophil-activating protein A and heat shock protein 10. Conclusion This experiment provides an impetus for further researches of the mechanism of H.pylori from spiral to dormant coccoid form, of development of new treatment strategy and vaccine design.