sGP Serves as a Structural Protein in Ebola Virus Infection
sGP Serves as a Structural Protein in Ebola Virus Infection
复制标题
DOI：
10.1093/infdis/jir313
复制
发表时间：
2011-11-01
影响因子：
6.4
通讯作者：
Kawaoka, Yoshihiro
中科院分区：
文献类型：
--
作者：
Iwasa, Ayaka;Shimojima, Masayuki;Kawaoka, Yoshihiro
方法.我们从表达质粒中表达了ZEBOV GP（1，2）、VP 40和NP蛋白以及sGP蛋白，并通过使用Western印迹检测了所得的病毒样颗粒。通过使用流式细胞术用识别GP（1，2）构象表位的KZ 52抗体分析表达GP（2）与GP（1）或sGP组合的细胞。用VSV delta G* 假型（其表达GFP报告基因而不是G蛋白）制备编码sGP和GP变体的假型病毒，以测试sGP对感染性的贡献。Western blot和流式细胞术分析表明存在共价连接的sGP-GP（2）分子。VSV delta G*（sGP + GP（2））和VSV delta G*（GP（1，2））感染Vero E6细胞并被KZ 52抗体中和。sGP过表达可降低VSV delta G*（GP（1，2））滴度。ZEBOV sGP可替代GP（1），形成sGP-GP（2）复合物并赋予感染性。我们的研究表明sGP作为一种结构蛋白具有新的作用。
Methods. We expressed ZEBOV GP(1,2), VP40, and NP proteins, together with sGP protein, from expression plasmids and examined the resultant virus-like particles by using Western blot. Cells expressing GP(2) in combination with either GP(1) or sGP were analyzed by using flow cytometry with the KZ52 antibody, which recognizes a GP(1,2) conformational epitope. A VSV pseudotype, VSV delta G*, which expresses a GFP reporter gene instead of the G protein, was used to produce pseudotyped viruses encoding sGP and variants of GP to test the contribution of sGP to infectivity.Results. Western blot and flow cytometric analyses suggested the existence of a covalently linked sGP-GP(2) molecule. VSV delta G*(sGP + GP(2)) and VSV delta G*(GP(1,2)) infected Vero E6 cells and were neutralized by the KZ52 antibody. Overexpression of sGP reduced the titer of VSV delta G*(GP(1,2)).Conclusions. ZEBOV sGP can substitute for GP(1), forming a sGP-GP(2) complex and conferring infectivity. Our studies suggest a novel role for sGP as a structural protein.