大肠杆菌抗氧化应激反应的蛋白质组学分析.doc

大肠杆菌抗氧化应激反应的蛋白质组学分析 摘 要 细胞代谢过程中，部分电子逃逸出氧化还原系统以氧分子作为电子受体，产生具有毒害作用的高能量活性氧分子（reactive oxygen species，ROS）包括超氧阴离子、单线态氧、、羟自由基脂质过氧化的中间产物。活性氧是一种广谱杀菌剂，是宿主抗菌作用的有效方式。但细菌在长期的进化发展出多种对策来抵抗宿主杀菌作用为检测与此相关的蛋白质，本研究主要通过正常及氧化应激条件下对大肠杆菌的培养，差速离心法提取膜蛋白SDS，基质辅助激光解吸电离飞行时间串联质谱(MALDI-TOF-MS/MS)进行鉴定分析。麦芽糖孔蛋。 关键词活性氧氧化应激反应；MALDI-TOF ABSTRACT During the metabolism in the cell, oxygen molecules as an important electron acceptor, Also accompanied by some electronic escape from the redox system, produce some high energy reactive oxygen species which has poisoned, including superoxide anion (O2-), singlet oxygen, hydrogen peroxide, hydroxyl radical, also includes the intermediate products of lipid metabolism. Reactive oxygen species (ROS) is a broad-spectrum fungicide. It is one of the effective way to host cell antimicrobial. However, bacteria in the long-term evolution development of a variety of strategies to resist the host immune bactericidal action. To detect which proteins associated with this system, in this study, mainly through training two group of the pathogenic E. coli, the one Treatment by hydrogen peroxide, the other one as the control group, Broken cells by ultrasound. Extraction the membrane proteins by differential centrifugation, make a SDSelectrophoresis with the Protein, Digging the significant difference protein band, after gel digestion, then identified by MALDI-TOF-MS/MS. We found two proteins which have significant difference, they are Maltoporin and outer membrane protein A (OmpA). Maltoporin and OmpA are associate to the small molecules penetrate into the cells. Therefore, we can speculate that to adapt to the environment Bacteria Close membrane channels of small molecules to reduce the intake of intracellular hydrogen peroxide, thereby reducing the toxicity of hydrogen peroxide. Through this experiment, we can initially clarify the oxidative stress mechanism of E. coli. Keywords: Reactive oxygen species; Oxidative stress; hydrogen peroxide; Membrane protein;