Abstract |
Fusion between fluorescently labeled plasma membrane vesicles (PMV) and cells expressing vesicular stomatitis virus (VSV) glycoprotein ( G-protein) was investigated by utilizing a lipid mixing assay based on fluorescence dequenching of octadecyl rhodamine (R18). The PMVs were prepared from Vero cells by hypotonic lysis. The G-protein was expressed on the cell surface either following infection with intact VSV or with an adenovirus vector (AdG12) containing the gene for the G-protein. Fusion was temperature and pH dependent and was inhibited by VSV G-antiserum. The pH dependence of PMV fusion paralleled that observed for VSV-cell fusion and VSV-induced syncytia formation. The kinetics of fusion followed an exponential dependence on time without an observable time lag after lowering pH. These findings indicate that dequenching R18-labeled PMV reliably represents the basic features of fusion of VSV with cells and can be used as a new tool in the study of fusion activity of virus envelope proteins expressed in cells.
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Authors | A Puri, M Krumbiegel, D Dimitrov, R Blumenthal |
Journal | Virology
(Virology)
Vol. 195
Issue 2
Pg. 855-8
(Aug 1993)
ISSN: 0042-6822 [Print] United States |
PMID | 8393251
(Publication Type: Journal Article)
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Chemical References |
- G protein, vesicular stomatitis virus
- Membrane Glycoproteins
- Rhodamines
- Viral Envelope Proteins
- octadecyl Rhodamine B chloride
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Topics |
- Animals
- Cell Membrane
(metabolism)
- Cloning, Molecular
- Fluorescence
- HeLa Cells
- Humans
- Hydrogen-Ion Concentration
- Membrane Fusion
- Membrane Glycoproteins
- Rhodamines
- Vero Cells
- Vesicular stomatitis Indiana virus
(metabolism)
- Viral Envelope Proteins
(metabolism)
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