Solubilization and reconstitution of vesicular stomatitis virus envelope using octylglucoside.
Reconstituted vesicular stomatitis virus envelopes or virosomes are formed by detergent removal from solubilized intact virus. We have monitored the solubilization process of the intact vesicular stomatitis virus by the nonionic surfactant octylglucoside at various initial virus concentrations by employing turbidity measurements. This allowed us to determine the phase boundaries between the membrane and the mixed micelles domains. We have also characterized the lipid and protein content of the solubilized material and of the reconstituted envelope. Both G and M proteins and all of the lipids of the envelope were extracted by octylglucoside and recovered in the reconstituted envelope. Fusion activity of the virosomes tested either on Vero cells or on liposomes showed kinetics and pH dependence similar to those of the intact virus.
Paternostre
M
M
Equipe Physicochimie des Systèmes Polyphasés, URA CNRS 1218, Université Paris Sud, Châtenay Malabry, France. paternos@cep.u-psud.fr
Viard
M
M
Meyer
O
O
Ghanam
M
M
Ollivon
M
M
Blumenthal
R
R
D000818:Animals; D002522:Chlorocebus aethiops; D003902:Detergents; D004591:Electrophoresis, Polyacrylamide Gel; D005453:Fluorescence; D005960:Glucosides; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008081:Liposomes; D008561:Membrane Fusion; D009391:Nephelometry and Turbidimetry; D019251:Nucleocapsid; D019590:Nucleocapsid Proteins; D010714:Phosphatidylethanolamines; D011137:Polystyrenes / Q000378:metabolism; D012235:Rhodamines; D012965:Sodium Chloride; D012995:Solubility; D014709:Vero Cells; D014721:Vesicular stomatitis Indiana virus / Q000737:chemistry* / Q000502:physiology; D014759:Viral Envelope Proteins / Q000737:chemistry*
D003902:Detergents; D005960:Glucosides; D008081:Liposomes; D019590:Nucleocapsid Proteins; D010714:Phosphatidylethanolamines; D011137:Polystyrenes; D012235:Rhodamines; D014759:Viral Envelope Proteins; C046703:lissamine-rhodamine-phosphatidylethanolamine; C018619:octyl-beta-D-glucoside; D012965:Sodium Chloride; C042077:N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)phosphatidylethanolamine; C003771:Amberlite XAD-2 resin