Endothelial adhesion molecule ESAM binds directly to the multidomain adaptor MAGI-1 and recruits it to cell contacts
Endothelial adhesion molecule ESAM binds directly to the multidomain adaptor MAGI-1 and recruits it to cell contacts
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DOI：
10.1016/j.yexcr.2004.07.010
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发表时间：
2004-10-15
影响因子：
3.7
通讯作者：
Butz, S
中科院分区：
文献类型：
--
作者：
Wegmann, F;Ebnet, K;Butz, S
内皮细胞选择性粘附分子（ESAM）是一种与内皮紧密连接（TJ）相关的免疫球蛋白样跨膜蛋白。基于酵母双杂交筛选，我们确定了膜相关鸟苷酸激酶蛋白MAGI-1是ESAM的细胞内结合伙伴。MAGI-1是一种多结构域接头蛋白，可与跨膜、细胞骨架和信号分子结合，并定位于上皮细胞的紧密连接。MAGI-1很可能通过PDZ结构域介导的相互作用与ESAM的c端序列结合。通过下拉实验证实了ESAM与MAGI-1之间的直接相互作用。这两种蛋白在转染的中国仓鼠卵巢（CHO）细胞中形成稳定的复合物，可进行免疫分离。我们发现MAGI-1与人脐静脉内皮细胞（HUVECs）和小鼠内皮细胞的细胞接触有关，在那里它与ESAM共定位。在CHO细胞中，MAGI-1在细胞接触处的募集需要ESAM的存在。因此，ESAM可能参与在内皮紧密连接处锚定MAGI-1。(C) 2004爱思唯尔公司版权所有。
Endothelial cell-selective adhesion molecule (ESAM) is an immunoglobulin-like transmembrane protein associated with endothelial tight junctions (TJ). Based on a yeast two-hybrid screen, we have identified the membrane-associated guanylate kinase protein MAGI-1 as an intracellular binding partner of ESAM. MAGI-1 is a multidomain adaptor protein, which binds to transmembrane, cytoskeletal, and signaling molecules, and has been localized to tight junctions in epithelial cells. MAGI-1 associates with the very C-terminal sequence of ESAM most likely through a PDZ domain-mediated interaction. The direct interaction between ESAM and MAGI-1 was confirmed by pull-down experiments. The two proteins formed stable complexes in transfected Chinese hamster ovary (CHO) cells, which could be immunoisolated. We found MAGI-1 to be associated with cell-cell contacts in human umbilical vein endothelial cells (HUVECs) and in mouse endothelium, where it colocalizes with ESAM. In CHO cells, recruitment of MAGI-1 to cell contacts required the presence of ESAM. Hence, ESAM may be involved in anchoring MAGI-1 at endothelial tight junctions. (C) 2004 Elsevier Inc. All rights reserved.