Zhang Yaning, Zou Yao, Ma Peipei, Muhammad Hassan Mushtaq, Li Yufeng, Jiang Ping
Key Laboratory of Bacteriology, Ministry of Agriculture, College of Veterinary Medicine, Nanjing Agricultural University, 210095, Nanjing, China.
Arch Microbiol. 2015 Mar;197(2):277-83. doi: 10.1007/s00203-014-1050-7. Epub 2014 Oct 26.
Adhesion protein MSG1 mediating adherence to porcine erythrocytes in Mycoplasma suis (M. suis) invasion has been identified previously. In order to determine the host membrane proteins that interact with MSG1, recombinant His-tagged MSG1 (rMSG1) was used to screen for interacting proteins in the protein extracts of porcine erythrocyte membrane. Potential rMSG1-interacting proteins were initially identified as band 3 and β-actin with molecular weight of 46 and 45 kDa, respectively. Immune fluorescence results showed that rMSG1 can specifically bind with the β-actin of HeLa, BHK-21, and HEK-293A cells, respectively. RNA interference assays further demonstrated that the interaction between β-actin and rMSG1 on HeLa cells was specific and dose dependent. Confocal microscopy showed that both rMSG1 and M. suis can partially co-localize with β-actin on the surface of porcine erythrocytes. Pull-down assays showed that rMSG1 can directly interact with β-actin. Our study is the first to report the interaction of MSG1 with β-actin, which will be of help to understand the pathogenesis of M. suis and develop a cultivation system.
黏附蛋白MSG1在猪支原体（M. suis）入侵过程中介导对猪红细胞的黏附，此前已被鉴定。为了确定与MSG1相互作用的宿主膜蛋白，使用重组His标签的MSG1（rMSG1）在猪红细胞膜蛋白提取物中筛选相互作用蛋白。最初鉴定出潜在的与rMSG1相互作用的蛋白为分子量分别为46 kDa和45 kDa的带3蛋白和β-肌动蛋白。免疫荧光结果表明，rMSG1可分别与HeLa、BHK-21和HEK-293A细胞的β-肌动蛋白特异性结合。RNA干扰试验进一步证明，HeLa细胞上β-肌动蛋白与rMSG1之间的相互作用具有特异性且呈剂量依赖性。共聚焦显微镜显示，rMSG1和猪支原体均可在猪红细胞表面与β-肌动蛋白部分共定位。下拉试验表明，rMSG1可直接与β-肌动蛋白相互作用。我们的研究首次报道了MSG1与β-肌动蛋白的相互作用，这将有助于了解猪支原体的发病机制并开发培养系统。
Zotero 插件