E-cadherin interactome complexity and robustness resolved by quantitative proteomics.
E-cadherin interactome complexity and robustness resolved by quantitative proteomics.
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DOI：
10.1126/scisignal.2005473
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发表时间：
2014-12-02
影响因子：
7.3
通讯作者：
Zaidel-Bar R
中科院分区：
文献类型：
--
作者：
Guo Z;Neilson LJ;Zhong H;Murray PS;Zanivan S;Zaidel-Bar R
E-钙粘蛋白介导的细胞间粘附和信号传导在健康上皮组织的发育和维持中起重要作用。粘附性由钙粘蛋白细胞外结构域赋予，并由与细胞质尾区相关的衔接子和酶的组装调节。在这里，我们采用邻近生物素化和定量蛋白质组学分离和鉴定612蛋白质附近的E-钙粘蛋白的胞质尾巴。我们使用蛋白质-蛋白质相互作用的结构信息数据库来构建迄今为止最全面的E-钙粘蛋白相互作用组，包含89个已知的E-钙粘蛋白体组分和346个新蛋白。此外，通过GFP标记的融合蛋白的克隆和表达，我们将26种新蛋白定位于粘附连接。最后，采用钙耗竭和肌球蛋白抑制，我们表明E-钙粘蛋白相互作用组是非常强大的扰动和基本上独立的细胞-细胞连接或肌动球蛋白收缩。
E-cadherin-mediated cell-cell adhesion and signaling plays an essential role in development and maintenance of healthy epithelial tissues. Adhesiveness is conferred by cadherin extracellular domains, and is regulated by an assembly of adaptors and enzymes associated with the cytoplasmic tail. Here, we employed proximity biotinylation and quantitative proteomics to isolate and identify 612 proteins in the vicinity of E-cadherin’s cytoplasmic tail. We used a structure-informed database of protein-protein interactions to construct the most comprehensive E-cadherin interactome to date, containing 89 known E-cadhesome components and 346 novel proteins. Moreover, through cloning and expression of GFP-tagged fusion proteins we localized 26 of the novel proteins to adherens junctions. Finally, employing calcium depletion and myosin inhibition we show the E-cadherin interactome to be remarkably robust to perturbation and essentially independent of cell-cell junctions or actomyosin contractility.