The interaction between Sendai virus and cell membranes: A quantitative analysis of 125I-Sendai virus particles' association with human red blood cells |
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Authors: | D. Wolf I. Kahana S. Nir A. Loyter |
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Affiliation: | Department of Biological Chemistry, The Hebrew University of Jerusalem, Institute of Life Sciences, Jerusalem, Israel |
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Abstract: | Intact Sendai virus particles were radiolabeled by the use of chloramine-T and Na 125I. The method described is reproducible, efficient and appropriate for the preparation of large quantities of biologically active virus with relatively high specific activity. Gel electrophoresis analysis of the radiolabeled virus revealed that approx. 50% of the total 125I incorporated in the virus are associated with the two viral envelope glycoproteins, while the remaining 50% are evenly distributed throughout the other viral polypeptides. The 125I-virus particles were used to study some of the kinetic parameters of the interaction between Sendai virus particles and human erythrocytes. Binding of virus particles at 4 °C is irreversible, non-cooperative and exhibits a characteristic saturation curve. A maximum of 1–2 × 103 virus particles bound per cell was derived from the saturation curve. Non-radioactive native virus particles as well as isolated glycophorin molecules competitively inhibit binding of the 125I-virus particles to human erythrocytes. Incubation at 37 °C of the virus-erythrocyte complex resulted in the release of about 33% of the bound virus to the surrounding medium. |
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