Andrews D
Department of Biochemistry, McMaster University, Hamilton, Ontario, Canada.
Biotechniques. 1989 Oct;7(9):960-2, 964-7.
A variety of assays have been developed which permit rapid and unambiguous determination of the membrane topology adopted by newly synthesized proteins. Cell-free systems and microinjected Xenopus oocytes are two of the most attractive approaches for characterizing the elements in both the nascent polypeptide and the membrane which together determine the final orientation of the protein in the membrane. Careful analysis of the mechanism of protein translocation using these methods has revealed a number of unusual topologies. The applications of a number of different assays for endoplasmic reticulum membrane translocation are described for the most commonly used cell-free systems (wheat germ and reticulocyte lysate), as well as for microinjected Xenopus oocytes.
已经开发出多种检测方法，可快速且明确地确定新合成蛋白质所采用的膜拓扑结构。无细胞系统和显微注射的非洲爪蟾卵母细胞是表征新生多肽和膜中共同决定蛋白质在膜中最终取向的元件的两种最具吸引力的方法。使用这些方法对蛋白质转运机制进行仔细分析，揭示了许多不同寻常的拓扑结构。本文描述了多种不同的内质网膜转运检测方法在最常用的无细胞系统（麦胚和网织红细胞裂解物）以及显微注射的非洲爪蟾卵母细胞中的应用。
