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Solubilization of the semliki forest virus membrane with sodium deoxycholate
Authors:Ari Helenius  Erik Fries  Henrik Garoff  Kai Simons
Affiliation:1. European Molecular Biology Laboratory, Postfach 10.2209, D-69 Heidelberg G.F.R.;2. Department of Serology and Bacteriology, University of Helsinki, Helsinki Finland
Abstract:The effects of increasing concentrations of sodium deoxycholate on Semliki Forest virus have been studied. Sodium deoxycholate begins to bind to the virus at less than 0.1 mM free equilibrium concentration and causes lysis of the viral membrane at 0.9 ± 0.1 mM free equilibrium concentration when 2.2 ± 0.2 ß 103mol of sodium deoxycholate are bound per mol of virus. Liberation of proteins from the membrane begins at 1.5 ± 0.1 mM sodium deoxycholate and the proteins released are virtually free from phospholipid above 2.0 mM sodium deoxycholate. The overall mechanism of sodium deoxycholate solubilization of the viral membrane resembles that of Triton X-100 and sodium dodecyl sulphate except that with sodium deoxycholate the various stages of membrane disruption occur at about 10-fold higher equilibrium free detergent concentrations. At sodium deoxycholate concentrations higher than 2.3 mM the viral spike glycoproteins can be separated by sucrose gradient centrifugation or gel filtration into constituent polypeptides E1, E2 and E3. E1 carries the haemagglutinating activity of the virus.
Keywords:SF virus  Semliki Forest virus
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