Watts C, Silver P, Wickner W
Cell. 1981 Aug;25(2):347-53. doi: 10.1016/0092-8674(81)90053-2.
The major coat protein of coliphage M13 is an integral protein of the E. coli plasma membrane prior to its assembly into new virus particles. It is generated from its precursor, procoat, by a membrane-bound leader peptidase. We now describe the reconstitution of a highly purified preparation of this enzyme into vesicles of E. coli phospholipids. These vesicles bind procoat made in vitro and procoat isolated from in vitro synthesis. Both the crude and the purified substrates were converted post-translationally to coat protein. A significant proportion of the coat protein becomes inserted into the vesicle bilayer, with the N terminus facing the vesicle interior and the C terminus exposed to the external medium. These results strongly suggest that highly purified leader peptidase from E. coli and phospholipids are the only components necessary to mediate the binding, processing and insertion of this integral membrane protein.
在装配成新的病毒颗粒之前，大肠杆菌噬菌体M13的主要外壳蛋白是大肠杆菌质膜的一种整合蛋白。它由其前体蛋白原衣壳通过膜结合的前导肽酶产生。我们现在描述了将这种酶的高度纯化制剂重构到大肠杆菌磷脂囊泡中的过程。这些囊泡结合体外合成的原衣壳以及从体外合成中分离出的原衣壳。粗底物和纯化底物都在翻译后转化为外壳蛋白。相当一部分外壳蛋白插入囊泡双层中，其N端朝向囊泡内部，C端暴露于外部介质。这些结果有力地表明，来自大肠杆菌的高度纯化的前导肽酶和磷脂是介导这种整合膜蛋白的结合、加工和插入所必需的唯一成分。
