Machamer C E, Doms R W, Bole D G, Helenius A, Rose J K
Department of Cell Biology, Yale University School of Medicine, New Haven, Connecticut 06510.
J Biol Chem. 1990 Apr 25;265(12):6879-83.
To investigate the function of heavy chain binding protein (BiP, GRP 78) in the endoplasmic reticulum, we have characterized its interaction with a model plasma membrane glycoprotein, the G protein of vesicular stomatitis virus. We used a panel of well characterized mutant G proteins and immunoprecipitation with anti-BiP antibodies to determine if BiP interacted with newly synthesized G protein and/or mutant G proteins retained in the endoplasmic reticulum. We made three major observations: 1) BiP bound transiently to folding intermediates of wild-type G protein which were incompletely disulfide-bonded; 2) BiP did not bind stably to all mutant G proteins which remain in the endoplasmic reticulum; and 3) BiP bound stably only to mutant G proteins which do not form correct intrachain disulfide bonds.
为了研究内质网中重链结合蛋白(BiP,GRP 78)的功能,我们对其与模型质膜糖蛋白——水疱性口炎病毒G蛋白的相互作用进行了表征。我们使用了一组特征明确的突变型G蛋白,并通过抗BiP抗体进行免疫沉淀,以确定BiP是否与新合成的G蛋白和/或滞留在内质网中的突变型G蛋白相互作用。我们有三个主要发现:1)BiP与野生型G蛋白的折叠中间体短暂结合,这些中间体的二硫键未完全形成;2)BiP并不稳定结合于所有滞留在内质网中的突变型G蛋白;3)BiP仅稳定结合于不能形成正确链内二硫键的突变型G蛋白。