凡纳滨对虾血蓝蛋白与病原菌凝集作用靶标的鉴定  

Identification of the Pathogen Agglutinative Target Recognized by Litopenaeus vannamei Hemocyanin

作　　者：章跃陵[1] 严芳[1] 樊大军[2] 蒋瑞萍[1] 胡忠[1] 李远友[3] 

机构地区：[1]汕头大学理学院生物学系,广东汕头515063 [2]汕头大学医学院第二附属医院,广东汕头515041 [3]汕头大学海洋生物研究所,广东汕头515063

出　　处：《中国生物化学与分子生物学报》2008年第5期457-462,共6页Chinese Journal of Biochemistry and Molecular Biology

基　　金：广东省自然科学基金项目(No.06027212);汕头市科技计划项目(No.2006-148);国家自然科学基金项目(No.30570325);广东省科技计划项目(No.2006A36502004)~~

摘　　要：血蓝蛋白是一种具有多种非特异性免疫学活性的多功能蛋白,以前的研究发现,血蓝蛋白具有凝集活性.本研究采用凝集抑制实验和亲和蛋白质组学等方法探索凡纳滨对虾血蓝蛋白与病原菌的凝集作用靶标.结果显示,大肠杆菌K12和副溶血弧菌外膜蛋白可以抑制血蓝蛋白对7种细菌的凝集活性,其中大肠杆菌K12中2种分子质量分别为16kD、18kD(命名为p16、p18)的外膜蛋白可以与血蓝蛋白发生特异性的结合,经MALDI-TOF/MS鉴定,p16、p18分别与大肠杆菌外膜蛋白OmpC、OmpX具有高度同源性.尤其是与大肠杆菌K12野生菌株相比,血蓝蛋白对ΔOmpX的凝集特异性明显降低,后者仅为前者的25%.由此推测,OmpX应为血蓝蛋白与病原菌的凝集作用靶标.Hemocyanin is a non-specific immune protein with multi-functions, has previous reported from our work. In this study, we used agglutination and its agglutinative activity inhibition assay and affinity proteomic technology to further investigate the agglutinative target of hemocyanin. The results have shown that the outer membrane proteins （Omps） of Escherichia coli K12 and Vibrio parahaemolyticus could completely inhibit the agglutinative activity of Litopenaeus vannamei hemocyanin against seven bacterial species, including E. coli K12, V. parahaemolyticus, Vibro alginolyticus, Vibro fluvialis, Vibro harveyi, Aeromonas hydrophila and Staphylococcus aureus. Two specific Omps with molecular weight of 16 kD and 18 kD （pl6 and p18） identified by MALDI-TOF/MS from E. coli K12 were able to bind hemocyanin, which were homologous with OmpC and OmpX of E. coli, respectively. The specific agglutinative activity of hemocyanin with AOmpX E. coli was substentially decreased by 75 % as comparison with wild type K12. We conclude that OmpX appeared to be the pathogen agglutinative target that recognized by hemocyanin.

关 键 词：血蓝蛋白 凝集活性 作用靶标 亲和蛋白质组学 外膜蛋白X 

分 类 号：Q513[生物学—生物化学]