The in vitro interaction of CmeA with CmeC  

作　　者：Hua-Wei Zhang Ximin Zeng Qi Qi Kai-Lei Sun Chong-Jun Ma Xiao-Jian Hu Jun Lin 

机构地区：[1]School of Life Sciences, Fudan University [2]Department of Animal Science, University of Tennessee, 2640 Morgan Circle Drive, Knoxville, TN 37996, USA

出　　处：《Chinese Science Bulletin》2014年第25期3114-3119,共6页

基　　金：supported by the National Basic Research Program of China(2009CB918602);the National Natural Science Foundation of China(30970575 and 31011120381);Shanghai Leading Academic Discipline Project(B111);Qi Qiwas supported by the National Talent Training Fund in Basic Research of China(J1210012)

摘　　要：The membrane fusion protein CmeA and the outer membrane channel CmeC,two important components of CmeABC system in Campylobacter jejuni,were expressed in Escherichia coli.After Ni-NTA and ion exchange columns purification,size exclusion chromatography showed that CmeA primarily existed as trimer and CmeC existed as monomer.Then the interaction between CmeA and CmeC was analyzed by dithiobis(succinimidyl propionate)(DSP)chemical crosslinking,pull-down assay on a Ni-NTA column,and isothermal titration calorimetry(ITC)measurement.The results clearly showed the CmeA–CmeC complex band,which confirmed the interaction in vitro and this interaction is independent of substrate and CmeB.It suggests that the mechanism underlying CmeABC function in C.jejuni is similar to that of AcrABTolC in E.coli.The membrane fusion protein CmeA and the outer membrane channel CmeC, two important compo- nents of CrneABC system in Campylobacter jejuni, were expressed in Escherichia coll. After Ni-NTA and ion exchange columns purification, size exclusion chromatography showed that CmeA primarily existed as trimer and CmeC existed as monomer. Then the interaction between CmeA and CmeC was analyzed by dithiobis （succinimidyl propionate） （DSP） chemical crosslinking, pull-down assay on a Ni-NTA column, and isothermal titration calorimetry （ITC） measurement. The results clearly showed the CmeACmeC complex band, which confirmed the interaction in vitro and this interaction is independent of substrate and CmeB. It suggests that the mechanism underlying CmeABC function in C. jejuni is similar to that of AcrAB TolC in E. coli.

关 键 词：相互作用 体外 等温滴定量热法 尺寸排阻色谱 大肠杆菌 膜融合蛋白 空肠弯曲菌 ABC系统 

分 类 号：R378[医药卫生—病原生物学]