The Ras recruitment system, a novel approach to the study of protein-protein interactions
The Ras recruitment system, a novel approach to the study of protein-protein interactions
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DOI：
10.1016/s0960-9822(98)70467-1
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发表时间：
1998-10-08
期刊：
影响因子：
9.2
通讯作者：
Aronheim, A
中科院分区：
文献类型：
--
作者：
Broder, YC;Katz, S;Aronheim, A
酵母双杂交系统是目前可用于识别和表征蛋白质相互作用的最有效的方法之一[1-4]。虽然这一过程非常强大，但仍存在一些问题和固有的局限性[5]。最近开发了一种新的系统，SOS招聘系统(SRS)，它基于与双混合系统不同的读数[6-8]。SRS克服了双杂交系统的一些局限性，因此是研究已知和新蛋白质之间蛋白质-蛋白质相互作用的一个有吸引力的替代方案。然而，我们在使用SRS时遇到了一些问题，因此我们开发了一种改进的蛋白质招募系统，称为RAS招募系统(RRS)，基于RAS定位于质膜以发挥其功能的绝对要求[9，10]。RAS膜的定位和激活可以通过两个杂合蛋白之间的相互作用来实现。我们已经使用五种不同的已知蛋白质-蛋白质相互作用证明了新型RRS系统的有效性，并通过文库筛选协议鉴定了两种以前未知的蛋白质-蛋白质相互作用。第一个相互作用(在这里详细描述)是基本亮氨酸拉链(BZIP)家族的成员JDP2和CCAAT/增强子结合蛋白(C/EBP)家族的成员C/EBP Gamma之间的相互作用。第二个相互作用是p21激活的蛋白激酶pak65和一个小G蛋白之间的相互作用(在Aronheim等人的随附论文中描述)。[11]))。RRS系统显著扩展了前面描述的SRS系统的有用性，并克服了它的几个限制。(C)现代生物有限公司ISSN 0960-9822。
The yeast two-hybrid system represents one of the most efficient approaches currently available for identifying and characterizing protein-protein interactions [1-4]. Although very powerful, this procedure exhibits several problems and inherent limitations [5]. A new system, the Sos recruitment system (SRS), was developed recently [6] based on a different readout from that of the two-hybrid system [6-8]. SRS overcomes several of the limitations of the two-hybrid system and thus serves as an attractive alternative for studying protein-protein interactions between known and novel proteins. Nevertheless, we encountered a number of problems using SRS and so have developed an improved protein recruitment system, designated the Ras recruitment system (RRS), based on the absolute requirement that Ras be localized to the plasma membrane for its function [9,10]. Ras membrane localization and activation can be achieved through interaction between two hybrid proteins. We have demonstrated the effectiveness of the novel RRS system using five different known protein-protein interactions and have identified two previously unknown protein-protein interactions through a library screening protocol. The first interaction (detailed here) is between JDP2, a member of the basic leucine zipper (bZIP) family, and C/EBP gamma, a member of the CCAAT/enhancer-binding protein (C/EBP) family. The second interaction is between the p21-activated protein kinase Pak65 and a small G protein (described in the accompanying paper by Aronheim et al. [11]). The RRS system significantly extends the usefulness of the previously described SRS system and overcomes several of its limitations. (C) Current Biology Ltd ISSN 0960-9822.