Paternostre M T, Lowy R J, Blumenthal R
Section on Membrane Structure and Function, LTB, NCI, Bethesda, MD 20892.
FEBS Lett. 1989 Jan 30;243(2):251-8. doi: 10.1016/0014-5793(89)80139-5.
Reconstituted vesicular stomatitis virus (VSV) envelopes were formed by solubilization of the viral envelope with Triton X-100 followed by removal of detergent by direct addition of SM2 biobeads. We provide direct demonstration of fusion of reconstituted VSV with cells using fluorescent lipid and aqueous probes incorporated into the VSV virosomes during reconstitution. We show a direct comparison of the kinetics and pH profile of fusion with cells between reconstituted VSV and fluorescently labeled intact virus. With this preparation it is now possible to gain additional information about the role of cooperativity in viral protein-mediated fusion, and to permit construction of efficient vehicles for delivery of drugs and other materials into cells.
重组水泡性口炎病毒(VSV)包膜是通过用Triton X-100溶解病毒包膜,然后直接添加SM2生物珠去除去污剂而形成的。我们利用重组过程中掺入VSV病毒体的荧光脂质和水性探针,直接证明了重组VSV与细胞的融合。我们展示了重组VSV与荧光标记的完整病毒在与细胞融合的动力学和pH曲线方面的直接比较。通过这种制备方法,现在有可能获得关于协同作用在病毒蛋白介导的融合中的作用的更多信息,并允许构建将药物和其他物质递送至细胞的有效载体。