Attachment of Sendai virus particles to cell membranes: dissociation of adsorbed virus particles with dithiothreitol.
Sendai virus particles bind to human erythrocytes at 4 degrees C and fuse with them at 37 degrees C. The present work describes a new method by which adsorbed virus particles can be removed from human erythrocytes, allowing quantitative determination of the number of virus particles which can bind and fuse with human erythrocyte membranes. Through the use of 125I-labeled Sendai virus particles, it is shown that incubation with 50 mM dithiothreitol removed about 90 to 95% of adsorbed virus particles. Fused virus particles were resistant to treatment with dithiothreitol. Negligible amounts of 125I-labeled Sendai virus particles were removed by treatment with dithiothreitol after incubation of virus-cell complexes at 37 degrees C. Trypsinized virus particles were able to attach to, but not fuse with, human erythrocytes even after prolonged incubation at 37 degrees C. Treatment with dithiothreitol removed as much as 80 to 85% of trypsinized virus particles incubated with human erythrocytes at 37 degrees C. A quantitative determination revealed that about 1,000 to 1,200 and 600 to 800 Sendai virus particles can bind to or fuse with human erythrocytes, respectively.
D000327:Adsorption; D004229:Dithiothreitol / Q000494:pharmacology*; D004910:Erythrocyte Membrane / Q000378:metabolism*; D006801:Humans; D010222:Parainfluenza Virus 1, Human / Q000378:metabolism*; D014357:Trypsin / Q000378:metabolism; D014771:Virion / Q000187:drug effects / Q000378:metabolism*
D014357:Trypsin; D004229:Dithiothreitol