Hug P, Sleight R G
Department of Molecular Genetics, Biochemistry, and Microbiology, University of Cincinnati College of Medicine, Ohio 45267-0524.
J Biol Chem. 1994 Feb 11;269(6):4050-6.
Virosomes were prepared by the insertion of vesicular stomatitis virus glycoprotein, a pH-sensitive fusion protein, into preformed liposomes. The fusogenic activity of these virosomes was characterized in cell-free fusion assays using liposomal targets. Fusion was monitored by concentration-dependent changes in the efficiency of resonance energy transfer between N-(lissamine rhodamine B sulfonyl)-phosphatidylethanolamine and N-(4-nitrobenzo-2-oxa-1,3-diazol)-phosphatidylethanolamine and by electron microscopy. The fusogenic activity was dependent on the presence of vesicular stomatitis virus glycoprotein, was pH-sensitive, and had a pH threshold of activation similar to that of the native virus. The extent of fusion was dependent upon the lipid composition of the vesicles. This technique will allow vesicles prepared by any method to be made fusogenic.
通过将水泡性口炎病毒糖蛋白（一种pH敏感的融合蛋白）插入预先形成的脂质体中来制备病毒体。在使用脂质体靶标的无细胞融合试验中对这些病毒体的融合活性进行了表征。通过监测N-（丽丝胺罗丹明B磺酰基）-磷脂酰乙醇胺和N-（4-硝基苯-2-恶唑-1,3-二氮唑）-磷脂酰乙醇胺之间共振能量转移效率的浓度依赖性变化以及通过电子显微镜来监测融合情况。融合活性取决于水泡性口炎病毒糖蛋白的存在，对pH敏感，并且具有与天然病毒相似的激活pH阈值。融合程度取决于囊泡的脂质组成。该技术将使通过任何方法制备的囊泡具有融合性。
