F protein induced fusion of Sendai viral envelopes with mouse teratocarcinoma cells through Le(x)-Le(x) interaction.
The efficiency of membrane fusion between reconstituted Sendai viral envelopes containing only the fusion protein (F-virosomes) and the plasma membrane of mouse teratocarcinoma cells (F9) in culture was assessed using an assay based on the relief of self-quenching of a lipid probe incorporated in the F-virosomes. The potential of F-virosomes was also evaluated for a targeted cytosolic delivery of lysozyme to F9 cells. [125I]Lysozyme entrapped into F-virosomes was taken to examine its fusion-mediated transfer to the F9 cells. Target specificity of the F-virosomes was confirmed by the interaction between the terminal Le(x) moiety (Gal beta 1-->4(Fuc alpha 1-->3)GlcNAc) of F protein and the Le(x) determinant on the membrane of F9 cells. Incubation of the loaded F-virosomes with cells led to fusion-mediated delivery, as inferred from the ability of cells to internalize lysozyme in the presence of azide (a potent inhibitor of endocytosis). These results suggest that carbohydrate-carbohydrate interaction is strong enough for target cell recognition followed by phospholipid bilayer melding induced by fusion glycoprotein of Sendai virus.
Kumar
M
M
Department of Biochemistry, University of Delhi, India.
Sarkar
D P
DP
D000818:Animals; D001692:Biological Transport; D002240:Carbohydrate Sequence; D002462:Cell Membrane / Q000502:physiology / Q000821:virology; D004705:Endocytosis; D007700:Kinetics; D016256:Lewis X Antigen / Q000378:metabolism*; D008561:Membrane Fusion; D051379:Mice; D008969:Molecular Sequence Data; D009113:Muramidase / Q000378:metabolism; D010222:Parainfluenza Virus 1, Human / Q000502:physiology*; D013347:Subcellular Fractions / Q000378:metabolism; D018243:Teratocarcinoma; D014407:Tumor Cells, Cultured; D014760:Viral Fusion Proteins / Q000378:metabolism*
D016256:Lewis X Antigen; D014760:Viral Fusion Proteins; D009113:Muramidase