Effect of Phospholipase C and Cholesterol Oxidase on Membrane Integrity, Microviscosity, and Infectivity of Vesicular Stomatitis Virus

Norman F. Moore*, Eric J. Patzer, Robert R. Wagner, Yechezkel Barenholz

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

66 Scopus citations

Abstract

Exposure of intact vesicular stomatitis (VS) virus or intact mixed-lipid vesicles to phospholipase-free cholesterol oxidase resulted in <5% oxidation of membrane cholesterol compared with >95% oxidation of cholesterol in detergentdisrupted virus or liposomes. Phospholipase C hydrolyzed ~55% of phospholipids in intact VS virion membranes and ~90% of phospholipids in small, single-walled lipid vesicles. Prior or simultaneous exposure of VS virions or liposomes to phospholipase C resulted in >90% oxidation by cholesterol oxidase of membrane cholesterol to cholest-4-en-3-one. Phospholipase A2 did not expose VS virion cholesterol to oxidation by cholesterol oxidase. Treatment with phospholipase C and cholesterol oxidase did not greatly alter the integrity of VS virion membrane, as determined by electron microscopy and protein electropherograms; only minimal amounts of oxidized cholesterol were released from virions exposed to both enzymes. Exposure to cholesterol oxidase alone resulted in only minor alterations in VS viral infectivity and membrane microviscosity monitored by fluorescence depolarization. Phospholipase C alone also had only a minimal effect on VS viral infectivity but markedly decreased the virion membrane apparent microviscosity. Treatment of VS virions with both phospholipase C and cholesterol oxidase caused a considerable decline in infectivity without further significant reduction in virion membrane microviscosity. Loss of VS viral infectivity appears to be due to the action of cholesterol oxidase on cholesterol exposed by the action of phospholipase C on phospholipid head groups in the outer surface of the viral membrane bilayer. This effect of cholesterol oxidase can be attributed to its oxidation of cholesterol in the viral membranes, or to production of hydrogen peroxide, or both.

Original languageEnglish
Pages (from-to)4708-4715
Number of pages8
JournalBiochemistry
Volume16
Issue number21
DOIs
StatePublished - 1 Oct 1977

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