Zhang J, Lautar S
Guilford Pharmaceuticals, Inc., Baltimore, Maryland 21224, USA.
Anal Biochem. 1996 Nov 1;242(1):68-72. doi: 10.1006/abio.1996.0429.
The yeast two-hybrid system is a powerful technique to detect binary protein interactions. In order to facilitate ternary complex analysis, we have developed the yeast three-hybrid system. Using epidermal growth factor (EGF) receptor, Grb2 and Sos proteins as an example, we demonstrated the use of the three-hybrid system to detect formation of a three-component complex, i.e., the Grb2-mediated interactions between EGF receptor and Sos. We then used the three-hybrid system to screen a library with Gal4-DNA binding domain -EGF receptor and Gal4 activation domain-Sos fusion proteins and isolated a plasmid encoding Grb2. Thus, the three-hybrid system is useful for characterizing and cloning components of ternary protein complexes.
酵母双杂交系统是检测二元蛋白质相互作用的一种强大技术。为便于进行三元复合物分析，我们开发了酵母三杂交系统。以表皮生长因子（EGF）受体、Grb2和Sos蛋白为例，我们展示了使用三杂交系统检测三元复合物的形成，即Grb2介导的EGF受体与Sos之间的相互作用。然后我们使用三杂交系统，以Gal4-DNA结合结构域-EGF受体和Gal4激活结构域-Sos融合蛋白筛选文库，并分离出一个编码Grb2的质粒。因此，三杂交系统对于表征和克隆三元蛋白质复合物的组分很有用。
