Active function of membrane receptors for enveloped viruses. I. Specific requirement for liposome-associated sialoglycolipids, but not sialoglycoproteins, to allow lysis of phospholipid vesicles by reconstituted Sendai viral envelopes.
Phospholipid liposomes composed of phosphatidylcholine (PC) and cholesterol (chol), bearing the sialoglycoprotein glycophorin (GP), are able to effectively bind Sendai virus particles, but not to be lysed by them. Incorporation of gangliosides (gangl) into the above phospholipid vesicles (yielding liposomes composed of PC/chol/gangl/GP), although not increasing their ability to interact with Sendai virions, rendered them susceptible to the viral lytic activity. This was inferred from the ability of the virus to induce release of carboxyfluorescein (CF) upon interaction at 37 degrees C with liposomes composed of PC/chol/gangl/GP. Lysis of liposomes required the presence of the two viral envelope glycoproteins, namely the hemagglutinin/neuraminidase (HN) and the fusion (F) polypeptides, and was inhibited by phenylmethyl sulfonylfluoride (PMSF), dithiothreitol (DTT) and trypsin, showing that virus-induced lysis of PC/chol/gangl/GP liposomes reflects the fusogenic activity of the virus. Incubation of Sendai virus particles with liposomes containing the acidic phospholipid dicetylphosphate (DCP) but lacking sialic acid containing receptors, also resulted in release of the liposome content. Lysis of these liposomes was due to the activity of the viral HN glycoprotein, therefore not reflecting the natural viral fusogenic activity. Fluorescence dequenching studies, using fluorescently labeled reconstituted Sendai virus envelopes (RSVE), have shown that the viral envelopes are able to fuse with neutral, almost to the same extent, as with negatively charged liposomes. However, fusion with negatively charged liposomes, as opposed to fusion with neutral liposomes, was mediated by the viral HN glycoprotein and not by the viral fusion polypeptide.
D002784:Cholesterol; D005732:Gangliosides; D006017:Glycolipids / Q000378:metabolism*; D006021:Glycophorins; D015707:HN Protein; D006387:Hemagglutination, Viral; D008081:Liposomes / Q000378:metabolism*; D010755:Organophosphates; D010222:Parainfluenza Virus 1, Human / Q000378:metabolism*; D010713:Phosphatidylcholines; D011991:Receptors, Virus / Q000378:metabolism*; D012795:Sialoglycoproteins / Q000378:metabolism; D014759:Viral Envelope Proteins / Q000378:metabolism; D014760:Viral Fusion Proteins
D005732:Gangliosides; D006017:Glycolipids; D006021:Glycophorins; D015707:HN Protein; D008081:Liposomes; D010755:Organophosphates; D010713:Phosphatidylcholines; D011991:Receptors, Virus; D012795:Sialoglycoproteins; D014759:Viral Envelope Proteins; D014760:Viral Fusion Proteins; C009255:sialoglycolipids; C002290:dicetylphosphate; D002784:Cholesterol