Ohlsson R I, Lane C D, Guengerich F P
Eur J Biochem. 1981 Apr;115(2):367-73. doi: 10.1111/j.1432-1033.1981.tb05247.x.
We described whole cell and cell-free systems capable of inserting into membranes cytochrome P-450 and epoxide hydratase made under the direction of rat liver RNA. The systems have been used to study the pathways followed by newly made secretory and integral membrane proteins. The cell-free system contains Xenopus laevis embryo membranes, and demonstrates competition for a common receptor between cytochrome P-450 and epoxide hydratase, and normal secretory proteins: evidence is provided for differential membrane receptor affinity. Thus, synthesis of secretory and membrane proteins appears to involve a common initial pathway. Microinjection of rat liver RNA into whole oocytes suggests that membrane insertion is neither cell type nor species specific, because functional rat liver enzymes are found inserted in the endoplasmic reticulum of the frog cell. Nonetheless, insertion is highly selective since albumin and several other proteins made under the direction of the injected liver RNA are sequestered within membrane vesicles and are then secreted by the oocyte, whilst epoxide hydratase and cytochrome P-450 are inserted into membranes but are not secreted.
我们描述了能够将在大鼠肝脏RNA指导下合成的细胞色素P - 450和环氧水解酶插入膜中的全细胞和无细胞系统。这些系统已被用于研究新合成的分泌蛋白和整合膜蛋白所遵循的途径。无细胞系统包含非洲爪蟾胚胎膜，并证明细胞色素P - 450、环氧水解酶与正常分泌蛋白之间对共同受体存在竞争：提供了膜受体亲和力差异的证据。因此，分泌蛋白和膜蛋白的合成似乎涉及共同的初始途径。将大鼠肝脏RNA显微注射到完整的卵母细胞中表明，膜插入既不具有细胞类型特异性也不具有物种特异性，因为在青蛙细胞的内质网中发现了功能性大鼠肝脏酶的插入。尽管如此，插入具有高度选择性，因为在注射的肝脏RNA指导下合成的白蛋白和其他几种蛋白质被隔离在膜泡内，然后由卵母细胞分泌，而环氧水解酶和细胞色素P - 450被插入膜中但不被分泌。
