Role of a transbilayer pH gradient in the membrane fusion activity of the influenza virus hemagglutinin: Use of the R18 assay to monitor membrane mergingReport as inadecuate




Role of a transbilayer pH gradient in the membrane fusion activity of the influenza virus hemagglutinin: Use of the R18 assay to monitor membrane merging - Download this document for free, or read online. Document in PDF available to download.

Biological Procedures Online

, Volume 1, Issue 3, pp 107–113

DOI: 10.1251-bpo13

Cite this article as: Ramalho-Santos, J. & de Lima, M.C.P. Biol Proced Online 1999 1: 107. doi:10.1251-bpo13

Abstract

It had been suggested that influenza virus-mediated membrane fusion might be dependent on a pH gradient across a target membrane. We have designed experiments in which this issue could be addressed. Two populations of liposomes were prepared, both simulating the plasma membrane of target cells, but with the pH of the internal aqueous medium buffered either at pH 7.4 physiological cytosol pH or at pH 5.0 endosomal pH at which influenza virus displays maximal fusion activity. By monitoring fusion using the R18 assay, we found that the internal pH of the target liposomes did not influence membrane merging as mediated by the influenza virus hemagglutinin, thus demonstrating that a transmembrane pH gradient is not required in this fusion process.

AbbreviationsHAinfluenza virus hemagglutinin

FCCPcarbonylcyanide-p-trifluoromethoxyphenyl hydrazone

LUVlarge unilamellar vesicles

PCphosphatidylcholine

PEphosphatidylethanolamine

R18octadecylrhodamine B chloride

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Author: João Ramalho-Santos - Maria C. Pedroso de Lima

Source: https://link.springer.com/







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