Expression and membrane integration of SARS-CoV E protein and its interaction with M protein
Expression and membrane integration of SARS-CoV E protein and its interaction with M protein
复制标题
DOI：
10.1007/s11262-009-0341-6
复制
发表时间：
2009-06-01
期刊：
影响因子：
1.6
通讯作者：
Li, Hui-Chun
中科院分区：
文献类型：
--
作者：
Chen, Shih-Chi;Lo, Shih-Yen;Li, Hui-Chun
克隆了严重急性呼吸综合征(SARS)冠状病毒E基因片段，并在体外和Vero E6细胞中以融合蛋白的形式表达。与其他N-糖基化蛋白类似，SARS-CoV E蛋白的糖基化是在微生物体存在的情况下进行共翻译的。SARS-CoV E蛋白被预测为一种双跨膜蛋白，缺乏传统的信号肽。这两个跨膜区(A.A.11-33和37-59)被预测为α-螺旋，它们自己穿透到膜中。不出所料，这两个跨膜区将一个细胞质蛋白插入内质网膜。这两个跨膜区中的任何一个与M蛋白共定位。E蛋白的两个跨膜区都需要与M蛋白相互作用，而亲水区(A.A.1-10或60-76)是可有可无的。这些结果对SARS-CoV组装的研究具有重要意义。
The severe acute respiratory syndrome (SARS)-CoV E gene fragment was cloned and expressed as a recombinant protein fused with a myc tag at the N-terminus in vitro and in Vero E6 cells. Similar to other N-glycosylated proteins, the glycosylation of SARS-CoV E protein occurred co-translationally in the presence of microsomes. The SARS-CoV E protein is predicted to be a double-spanning membrane protein lacking a conventional signal peptide. Both of the transmembrane regions (a.a. 11-33 and 37-59) are predicted to be alpha-helices, which penetrate into membranes by themselves. As expected, these two transmembrane regions inserted a cytoplasmic protein into the endoplasmic reticulum membrane. Either of these two transmembrane domains co-localized with M protein. Both the transmembrane domains of E protein are required to interact with M protein, while either of the hydrophilic regions (a.a. 1-10 or 60-76) is dispensable as shown by co-immunoprecipitation assay. These results are important for the study of SARS-CoV assembly.