Akt phosphorylates connexin43 on Ser373, a "Mode-1" binding site for 14-3-3
Akt phosphorylates connexin43 on Ser373, a "Mode-1" binding site for 14-3-3
复制标题
DOI：
10.1080/15419060701755958
复制
发表时间：
2007-09-01
影响因子：
--
通讯作者：
Warn-Cramer, Bonnie J.
中科院分区：
文献类型：
--
作者：
Park, Darren J.;Wallick, Christopher J.;Warn-Cramer, Bonnie J.
连接蛋白43（Cx43）是一种跨膜蛋白，其形成桥接相邻细胞之间的差距的通道，这允许细胞间的信息交换。Cx43受磷酸化和相互作用蛋白质的调节。与14-3-3的“模式-1”相互作用需要Cx43上Ser 373的磷酸化（Park等人，2006）。Akt磷酸化并靶向许多蛋白质以与14-3-3相互作用。在这里，我们证明了Akt磷酸化Ser 373和Ser 369上的Cx43;识别Akt磷酸化位点或磷酸化Ser 14-3-3结合位点的抗体识别来自EGF处理的细胞的蛋白质，该蛋白质作为Cx43迁移，GST-14-3-3结合EGF处理的细胞中内源性磷酸化的Cx43。共聚焦显微镜支持Cx43与Akt和14-3-3在间隙连接斑块外缘的共定位。这些数据表明，Akt可以靶向Cx43与14-3-3的相互作用，这可能在Cx43多聚体的前向运输和/或其并入现有的间隙连接斑块中发挥作用。
Connexin43 (Cx43) is a membrane-spanning protein that forms channels that bridge the gap between adjacent cells and this allows for the intercellular exchange of information. Cx43 is regulated by phosphorylation and by interacting proteins. "Mode-1" interaction with 14-3-3 requires phosphorylation of Ser373 on Cx43 (Park et al. 2006). Akt phosphorylates and targets a number of proteins to interactions with 14-3-3. Here we demonstrate that Akt phosphorylates Cx43 on Ser373 and Ser369; antibodies recognizing Akt-phosphorylated sites or phospho-Ser "rnode-1" 14-3-3-binding sites recognize a protein from EGF-treated cells that migrates as Cx43, and GST-14-3-3 binds to Cx43 phosphorylated endogenously in EGF-treated cells. Confocal microscopy supports the co-localization of Cx43 with Akt and with 14-3-3 at the outer edges of gap junctional plaques. These data suggest that Akt could target Cx43 to an interaction with 14-3-3 that may play a role in the forward trafficking of Cx43 multimers and/or their incorporation into existing gap junctional plaques.