Protein recruitment systems for the analysis of protein±protein interactions
Protein recruitment systems for the analysis of protein±protein interactions
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DOI：
10.1006/meth.2001.1153
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发表时间：
2001-05-01
期刊：
影响因子：
4.8
通讯作者：
Aronheim, A
中科院分区：
文献类型：
--
作者：
Aronheim, A
酿酒酵母是分析蛋白质+/-蛋白质相互作用的优秀遗传工具。迄今为止最常见的系统是双混合系统。尽管事实证明非常强大，但两种混合系统也存在一些固有的问题和局限性。最近，已经描述了两种替代系统，它们利用了这样一个事实：信号转导效应器定位于质膜内叶对于酵母的生存能力是绝对必要的。这些效应子可以是 Ras 鸟苷酸交换因子或 Ras 本身。两个系统中使用的酵母菌株都是酵母 Ras 鸟苷酸交换因子 CDC25 的温度敏感突变体。这些效应物的膜定位是通过蛋白质+/-蛋白质相互作用实现的。每个系统均可用于测试已知蛋白质对之间的相互作用，以及用于分离新的蛋白质相互作用。这里描述的是两种蛋白质招募系统要考虑的科学和技术步骤。 (C) 2001 年学术出版社。
The yeast Saccharomyces cerevisiae serves as an excellent genetic tool for the analysis of protein +/- protein interactions. The most common system, used to date, is the two-hybrid system. Although proven very powerful, the two-hybrid system exhibits several inherent problems and limitations. Recently, two alternative systems have been described that take advantage of the fact that localization of signal transduction effecters to the inner leaflet of the plasma membrane Is absolutely necessary for yeast viability. These effecters can either be the Ras guanyl nucleotide exchange factor or Ras itself. The yeast strain used in both systems is a temperature-sensitive mutant in the yeast Ras guanyl nucleotide exchange factor, CDC25. Membrane localization of these effecters is achieved via protein +/- protein interaction. Each system can be used to test interaction between known protein pairs, as well as for isolation of novel protein interactions. Described here are the scientific and technical steps to be considered for both protein recruitment systems. (C) 2001 Academic Press.