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Knock-in AP-MS: a sensitive platform for mapping protein interaction networks in the Wnt signaling pathway

机译:敲入AP-MS:用于在WNT信号通路中映射蛋白质交互网络的敏感平台

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Affinity-purification mass spectrometry (AP-MS) provides a sensitive method for the identification of protein complexes and characterization of protein networks and is ideally suited to the study of the Wnt signaling pathway in which communication between members of protein complexes is an important regulator of pathway activity. Knock-in techniques enable tagging of endogenous genes by introduction of epitope tags directly into the genome (Figure 1). These endogenously tagged genes can then be used as baits in AP-MS experiments, providing a sensitive platform for protein complex purification and identification. Two key players in the Wnt pathway (Figure 2), Adenomatous polyposis coli (APC) and beta-catenin (CTNNB1) are used as baits in this study to show the efficacy of the approach and potential for identification of novel interactors.
机译:亲和纯化质谱法(AP-MS)提供了鉴定蛋白质复合物的敏感方法和蛋白质网络的表征,并且理想地适用于WNT信号传导途径的研究,其中蛋白质复合物成员之间的通信是一个重要的调节器途径活动。通过将表位标签直接引入基因组(图1),敲入技术通过将表位标签引入到基因组中来说然后,这些内源标记的基因可以用作AP-MS实验中的诱饵,为蛋白质复合物纯化和鉴定提供敏感的平台。 WNT途径中的两个关键球员(图2),腺瘤性息肉组织大肠杆菌(APC)和β-连环蛋白(CTNNB1)被用作本研究中的诱饵,以显示方法的效果和鉴定新型交互运动的潜力。

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