pH-dependent fusion of reconstituted vesicular stomatitis virus envelopes with Vero cells. Measurement by dequenching of fluorescence.
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.
Paternostre
M T
MT
Section on Membrane Structure and Function, LTB, NCI, Bethesda, MD 20892.
Lowy
R J
RJ
Blumenthal
R
R
D000818:Animals; D003902:Detergents; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008561:Membrane Fusion; D008856:Microscopy, Fluorescence; D012995:Solubility; D013050:Spectrometry, Fluorescence; D014709:Vero Cells; D014721:Vesicular stomatitis Indiana virus / Q000378:metabolism; D014759:Viral Envelope Proteins / Q000378:metabolism*
D003902:Detergents; D014759:Viral Envelope Proteins