Influence of the Membrane Glycoprotein and Cholesterol of Vesicular Stomatitis Virus on the Dynamics of Viral and Model Membranes: Fluorescence Studies

Ranajit Pal, Jon R. Wiener, Yechezkel Barenholz, Robert R. Wagner*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

15 Scopus citations

Abstract

The interaction of the integral membrane glycoprotein of vesicular stomatitis virus with the phospholipid bilayer in reconstituted lipid vesicles was determined by differential polarized phase fluorometry, steady-state fluorescence anisotropy, and emission lifetime measurements. The glycoprotein was isolated from the viral membrane by solubilization with octyl β-glucopyranoside and was reconstituted into lipid vesicles by the detergent dialysis method. The fluorescent probes 1,6-diphenyl-1,3,5-hexatriene and 1-[4-(trimethylammonio)phenyl]-6-phenyl-1,3,5-hexatriene revealed that, in dipalmitoylphosphatidylcholine vesicles, the glycoprotein disorders the bilayer in the gel phase and orders it to a small extent in the liquid-crystalline phase. When the viral glycoprotein was reconstituted into 1-palmitoyl-2-oleoylphosphatidylcholine vesicles, only the ordering effect of the protein on the lipid bilayer was observed throughout the temperature range studied. The lifetime of both probes was found to be slightly shortened in the presence of the protein. No large effect of glycoprotein on the dynamics of the lipid phase was observed when the protein was reconstituted into total viral lipid vesicles. The role that cholesterol plays in determining the structural order of the viral membrane was determined by steady-state fluorescence anisotropy and differential polarized phase fluorometry. In intact viral membranes, as well as in viral lipid vesicles, the depolarization rotations of both 1,6-diphenyl-1,3,5-hexatriene and 1-[4-(trimethylammonio)-phenyl]-6-phenyl-1,3,5-hexatriene were highly hindered. Removal of cholesterol from the viral membrane, or from total viral lipid vesicles, disordered the membrane matrix markedly. Steady-state fluorescence anisotropy of trans-parinaric acid embedded in the viral envelope showed a broad phase separation in the viral membrane following depletion of cholesterol from the viral envelope. A similar effect was noted in cholesterol-free viral lipid vesicles. The results presented in this paper therefore demonstrate that both the membrane glycoprotein and cholesterol of vesicular stomatitis virus affect the dynamics of lipid bilayers to a significant extent. The effect of cholesterol was found to be greater than that of the protein. The role of cholesterol in determining the organization of glycoprotein spikes in the membrane of the virus and its implication for viral infectivity are discussed in light of these results.

Original languageEnglish
Pages (from-to)3624-3630
Number of pages7
JournalBiochemistry
Volume22
Issue number15
DOIs
StatePublished - 1983

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