Helenius A, Fries E, Kartenbeck J
J Cell Biol. 1977 Dec;75(3):866-80. doi: 10.1083/jcb.75.3.866.
The spike glycoproteins of the Semliki forest virus membrane have been incorporated into vesicular phospholipid bilayers by a detergent-dialysis method. The detergent used was beta-D-octylglucoside which is nonionic and has an exceptionally high critical micellar concentration which facilitates rapid removal by dialysis. The vesicles obtained were of varying sizes and had spikes on their surface. Two classes of vesicles were preferentially formed, small protein-rich and large lipid-rich (average lipid to protein weight ratios, 0.22 and 3.5, respectively). Both classes of vesicles retained the hemagglutinating activity of the virus. The proteins were attached to the lipid bilayer by hydrophobic peptide segments, as in the viral membrane. Most of the proteins were accessible to proteolytic digestion from the outside, suggesting an asymmetric orientation.
通过去污剂透析法,将塞姆利基森林病毒膜的刺突糖蛋白整合到囊泡磷脂双层中。所使用的去污剂是β-D-辛基葡糖苷,它是非离子型的,具有极高的临界胶束浓度,这有利于通过透析快速去除。得到的囊泡大小各异,表面有刺突。优先形成两类囊泡,富含蛋白质的小囊泡和富含脂质的大囊泡(平均脂质与蛋白质重量比分别为0.22和3.5)。两类囊泡都保留了病毒的血凝活性。蛋白质通过疏水肽段附着在脂质双层上,如同在病毒膜中一样。大多数蛋白质可从外部被蛋白酶消化,表明其具有不对称的取向。