The junction-associated protein AF-6 interacts and clusters with specific Eph receptor tyrosine kinases at specialized sites of cell-cell contact in the brain.
The junction-associated protein AF-6 interacts and clusters with specific Eph receptor tyrosine kinases at specialized sites of cell-cell contact in the brain.
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连接相关的蛋白AF-6在大脑中细胞细胞接触的专用部位与特定的EPH受体酪氨酸激酶相互作用。
DOI：
10.1083/jcb.144.2.361
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发表时间：
1999-01-25
影响因子：
7.8
通讯作者：
Hovens, C M
中科院分区：
文献类型：
--
作者：
Buchert, M;Schneider, S;Meskenaite, V;Adams, M T;Canaani, E;Baechi, T;Moelling, K;Hovens, C M
AF-6/afadin蛋白含有一个单一的PDZ结构域，在细胞-细胞连接的特殊部位形成细胞膜的外周成分。为了确定AF-6的潜在受体结合靶点，我们从具有潜在PDZ结构域结合末端的受体中选择一系列cooh末端肽筛选AF-6的PDZ结构域。AF-6的PDZ结构域通过其COOH末端在体外和体内与rtk的Eph亚家族的一部分成员相互作用。将绿色荧光蛋白标记的AF-6融合蛋白与全长Eph受体共转染到异源细胞中，可诱导Eph受体和AF-6在细胞-细胞接触部位聚集。成年大鼠脑免疫组织化学分析显示AF-6与Eph受体在海马兴奋性突触的突触后膜位点共聚。此外，AF-6是Eph受体亚群的底物，AF-6的磷酸化依赖于受体的功能性激酶结构域。内源性AF-6与Eph受体的物理相互作用通过全脑裂解物的共免疫沉淀得到证实。AF-6是在成年大鼠脑突触后特化中介导Eph受体聚集的候选物质。
The AF-6/afadin protein, which contains a single PDZ domain, forms a peripheral component of cell membranes at specialized sites of cell–cell junctions. To identify potential receptor-binding targets of AF-6 we screened the PDZ domain of AF-6 against a range of COOH-terminal peptides selected from receptors having potential PDZ domain-binding termini. The PDZ domain of AF-6 interacts with a subset of members of the Eph subfamily of RTKs via its COOH terminus both in vitro and in vivo. Cotransfection of a green fluorescent protein-tagged AF-6 fusion protein with full-length Eph receptors into heterologous cells induces a clustering of the Eph receptors and AF-6 at sites of cell–cell contact. Immunohistochemical analysis in the adult rat brain reveals coclustering of AF-6 with Eph receptors at postsynaptic membrane sites of excitatory synapses in the hippocampus. Furthermore, AF-6 is a substrate for a subgroup of Eph receptors and phosphorylation of AF-6 is dependent on a functional kinase domain of the receptor. The physical interaction of endogenous AF-6 with Eph receptors is demonstrated by coimmunoprecipitation from whole rat brain lysates. AF-6 is a candidate for mediating the clustering of Eph receptors at postsynaptic specializations in the adult rat brain.