Stagljar I, Korostensky C, Johnsson N, te Heesen S
Institute of Veterinary Biochemistry, University Zürich-Irchel, Winterthurerstrasse 190, CH-8057 Zurich, Switzerland.
Proc Natl Acad Sci U S A. 1998 Apr 28;95(9):5187-92. doi: 10.1073/pnas.95.9.5187.
A detection system for interactions between membrane proteins in vivo is described. The system is based on split-ubiquitin [Johnsson, N. & Varshavsky, A. (1994) Proc. Natl. Acad. Sci. USA 91, 10340-10344]. Interaction between two membrane proteins is detected by proteolytic cleavage of a protein fusion. The cleavage releases a transcription factor, which activates reporter genes in the nucleus. As a result, interaction between membrane proteins can be analyzed by the means of a colorimetric assay. We use membrane proteins of the endoplasmic reticulum as a model system. Wbp1p and Ost1p are both subunits of the oligosaccharyl transferase membrane protein complex. The Alg5 protein also localizes to the membrane of the endoplasmic reticulum, but does not interact with the oligosaccharyltransferase. Specific interactions are detected between Wbp1p and Ost1p, but not between Wbp1p and Alg5p. The new system might be useful as a genetic and biochemical tool for the analysis of interactions between membrane proteins in vivo.
本文描述了一种用于检测体内膜蛋白之间相互作用的检测系统。该系统基于分裂泛素[约翰松，N. & 瓦尔沙夫斯基，A.（1994年）美国国家科学院院刊91，10340 - 10344]。通过蛋白质融合的蛋白水解切割来检测两种膜蛋白之间的相互作用。切割释放出一种转录因子，该转录因子激活细胞核中的报告基因。因此，可以通过比色测定法来分析膜蛋白之间的相互作用。我们以内质网的膜蛋白作为模型系统。Wbp1p和Ost1p都是寡糖基转移酶膜蛋白复合物的亚基。Alg5蛋白也定位于内质网的膜上，但不与寡糖基转移酶相互作用。在Wbp1p和Ost1p之间检测到了特异性相互作用，但在Wbp1p和Alg5p之间未检测到。这种新系统可能作为一种遗传和生化工具，用于分析体内膜蛋白之间的相互作用。
