Automated yeast two-hybrid screening for nuclear receptor-interacting proteins
Automated yeast two-hybrid screening for nuclear receptor-interacting proteins
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DOI：
10.1074/mcp.m400169-mcp200
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发表时间：
2005-02-01
影响因子：
7
通讯作者：
Koegl, M
中科院分区：
文献类型：
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作者：
Albers, M;Kranz, H;Koegl, M
蛋白质-蛋白质相互作用的高通量分析是假设生成研究的一个重要领域。利用酵母双杂交系统的改进和自动化版本，我们完成了一个大型的相互作用筛选项目，重点是核受体及其辅助因子。425个独立的酵母双杂交cDNA文库筛选得到6425个潜在的相互作用蛋白片段，涉及1613个不同的相互作用对。我们表明，可以使用简单的统计参数将数据集缩小到377对交互对的高置信度集，其中经过验证的交互对丰富到所有对的61%。在高置信度范围内，有64种新的蛋白质可能与核受体或其辅助因子结合。我们讨论了几个高兴趣的例子，我们期望这个庞大的数据集的交流将有助于补充我们对这个转录因子家族的蛋白质相互作用库的知识，并激发各种新的候选相互作用物的表征。
High throughput analysis of protein-protein interactions is an important sector of hypothesis-generating research. Using an improved and automated version of the yeast two-hybrid system, we completed a large interaction screening project with a focus on nuclear receptors and their cofactors. A total of 425 independent yeast two-hybrid cDNA library screens resulted in 6425 potential interacting protein fragments involved in 1613 different interaction pairs. We show that simple statistical parameters can be used to narrow down the data set to a high confidence set of 377 interaction pairs where validated interactions are enriched to 61% of all pairs. Within the high confidence set, there are 64 novel proteins potentially binding to nuclear receptors or their cofactors. We discuss several examples of high interest, and we expect that communication of this huge data set will help to complement our knowledge of the protein interaction repertoire of this family of transcription factors and instigate the characterization of the various novel candidate interactors.