Use of virus-attached antibodies or insulin molecules to mediate fusion between Sendai virus envelopes and neuraminidase-treated cells.
Anti-human erythrocyte antibodies or insulin molecules were covalently coupled to the glycoproteins (the hemagglutinin/neuraminidase and the fusion polypeptides) of Sendai virus envelopes with N-succinimidyl 3-(2-pyridyldithio)propionate and succinimidyl 4-(p-maleimidophenyl)butyrate as cross-linking reagents. Reconstituted Sendai virus envelopes, bearing covalently attached anti-human erythrocyte antibodies or insulin molecules, were able to bind to but not fuse with virus receptor depleted human erythrocytes (neuraminidase-treated human erythrocytes). Only coreconstitution of Sendai virus glycoproteins, bearing attached anti-human erythrocyte antibodies or insulin molecules with intact, untreated viral glycoproteins, led to the formation of fusogenic, targeted reconstituted Sendai virus envelopes. Binding and fusion of reconstituted Sendai virus envelopes, bearing anti-human erythrocyte antibodies or insulin molecules, with neuraminidase-treated human erythrocytes were blocked by the monovalent fraction, obtained after papain digestion of immunoglobulins, made of anti-human erythrocyte antibodies or free insulin molecules, respectively. The results of this work demonstrate an active role of the viral binding protein (hemagglutinin/neuraminidase polypeptide) in the virus membrane fusion process and show a novel and efficient method for the construction of targeted, fusogenic Sendai virus envelopes.
Gitman
A G
AG
Kahane
I
I
Loyter
A
A
D000906:Antibodies; D004229:Dithiothreitol / Q000494:pharmacology; D004912:Erythrocytes / Q000276:immunology; D006023:Glycoproteins / Q000378:metabolism*; D006801:Humans; D007328:Insulin / Q000494:pharmacology*; D007700:Kinetics; D009439:Neuraminidase / Q000494:pharmacology*; D010222:Parainfluenza Virus 1, Human / Q000378:metabolism*; D011991:Receptors, Virus / Q000187:drug effects / Q000378:metabolism*; D014759:Viral Envelope Proteins / Q000378:metabolism*
D000906:Antibodies; D006023:Glycoproteins; D007328:Insulin; D011991:Receptors, Virus; D014759:Viral Envelope Proteins; D009439:Neuraminidase; D004229:Dithiothreitol