Paternostre M, Viard M, Meyer O, Ghanam M, Ollivon M, Blumenthal R
Equipe Physicochimie des Systèmes Polyphasés, URA CNRS 1218, Université Paris Sud, Châtenay Malabry, France.
Biophys J. 1997 Apr;72(4):1683-94. doi: 10.1016/S0006-3495(97)78814-3.
Reconstituted vesicular stomatitis virus envelopes or virosomes are formed by detergent removal from solubilized intact virus. We have monitored the solubilization process of the intact vesicular stomatitis virus by the nonionic surfactant octylglucoside at various initial virus concentrations by employing turbidity measurements. This allowed us to determine the phase boundaries between the membrane and the mixed micelles domains. We have also characterized the lipid and protein content of the solubilized material and of the reconstituted envelope. Both G and M proteins and all of the lipids of the envelope were extracted by octylglucoside and recovered in the reconstituted envelope. Fusion activity of the virosomes tested either on Vero cells or on liposomes showed kinetics and pH dependence similar to those of the intact virus.
重组水泡性口炎病毒包膜或病毒体是通过从溶解的完整病毒中去除去污剂形成的。我们通过使用浊度测量法,在不同初始病毒浓度下,用非离子表面活性剂辛基葡糖苷监测了完整水泡性口炎病毒的溶解过程。这使我们能够确定膜和混合胶束域之间的相界。我们还对溶解材料和重组包膜的脂质和蛋白质含量进行了表征。G蛋白和M蛋白以及包膜的所有脂质都被辛基葡糖苷提取,并在重组包膜中回收。在Vero细胞或脂质体上测试的病毒体的融合活性显示出与完整病毒相似的动力学和pH依赖性。