Protein interaction screening by quantitative immunoprecipitation combined with knockdown (QUICK)
Protein interaction screening by quantitative immunoprecipitation combined with knockdown (QUICK)
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DOI：
10.1038/nmeth972
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发表时间：
2006-12-01
期刊：
影响因子：
48
通讯作者：
Mann, Matthias
中科院分区：
文献类型：
--
作者：
Selbach, Matthias;Mann, Matthias
目前蛋白质-蛋白质相互作用（PPI）的筛选方法依赖于人工融合蛋白的过表达，使得难以评估体内相关性。在这里，我们结合联合收割机稳定同位素标记与细胞培养中的氨基酸（SILAC），RNA干扰（RNAi），免疫共沉淀和定量质谱分析，以非常高的置信度检测哺乳动物细胞中内源性蛋白质的细胞相互作用伙伴。我们使用该筛选来鉴定β-连环蛋白和Cbl的相互作用伴侣。
Present screening methods for protein-protein interactions (PPIs) rely on the overexpression of artificial fusion proteins, making it difficult to assess in vivo relevance. Here we combine stable isotope labeling with amino acids in cell culture (SILAC), RNA interference (RNAi), coimmunoprecipitation and quantitative mass-spectrometry analysis to detect cellular interaction partners of endogenous proteins in mammalian cells with very high confidence. We used this screen to identify interaction partners of beta-catenin and Cbl.