摘要
黏附因子在病原菌的致病过程中发挥了重要作用,鉴定新的黏附因子是了解病原菌致病机制的重要手段。本研究中首次发现游离肝素能够竞争性抑制副溶血弧菌与Hela细胞的黏附,表明细胞外基质中的肝素可能是细菌重要的细胞表面黏附受体。进一步利用肝素亲和层析技术垂钓到6种副溶血弧菌的外膜蛋白,并通过基因克隆和原核表达技术成功获得重组的外膜蛋白并对其进行了深入研究。重组蛋白IMPDH、EF-Tu和Opp A可黏附Hela细胞,并可以显著抑制副溶血弧菌与Hela细胞的黏附,说明这3种蛋白可能是其重要的潜在黏附因子。
Adhesins are critical to the infectious pathogens, thus identification of novel adhesins is very crucial to understanding the pathogen＇s infection mechanism. In this study, we demonstrated that heparin can competitively inhibits the adhesion of Vibrio parahaemolyticus（VP） to Hela cells, which indicates heparin in ECM could be the cell receptor of VP. Using heparin-sepharose affinity chromatography, six heparin-binding VP out-membrane proteins（OMPs） are selected. The six corresponding recombinant proteins were expressed and purified. According to the result of function verification test, recombinant protein IMPDHEF-Tu and Opp A are capable of binding to the Hela cells surface. Furthermore, they can inhibit the adhesion of VP to Hela cells significantly. Our research shows that the three OMPs are candidates of VP adhesins.
作者
杨娅楠
王婧婷
陈萌
耿绪云
袁增智
孙金生
YANG Yanan;WANG Jingting;CHEN Meng;GENG Xuyun;YUAN Zengzhi;SUN Jinsheng(Tianjin Key Laboratory of Animal and Plant Resistance, College of Life Science, Tianjin Normal University, Tianjin 300387, China;Tianjin Center for Control and Prevention of Aquatic Animal Infectious Disease, Tianjin 300221, China)
出处
《水产学报》
CAS
CSCD
北大核心
2017年第11期1779-1787,共9页
Journal of Fisheries of China
基金
国家“九七三”重点基础研究发展计划(2012CB114405)
国家自然科学基金(31402322)
天津市应用基础计划项目(15JCQNJC46400)
天津师范大学博士基金(52XB1412)
关键词
副溶血弧菌
肝素
外膜蛋白
黏附因子
Vibrio parahaemolyticus
heparin
out-membrane proteins
adhesins
