Regulation of Vascular Endothelial Growth Factor-induced Endothelial Cell Migration by LIM Kinase 1-mediated Phosphorylation of Annexin 1
Regulation of Vascular Endothelial Growth Factor-induced Endothelial Cell Migration by LIM Kinase 1-mediated Phosphorylation of Annexin 1
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DOI：
10.1074/jbc.m109.098665
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发表时间：
2010-03-12
影响因子：
4.8
通讯作者：
Huot, Jacques
中科院分区：
文献类型：
--
作者：
Cote, Maxime C.;Lavoie, Jessie R.;Huot, Jacques
在这项研究中，我们获得的证据表明，膜联蛋白1是p38/MAPKAP激酶-2途径的一个新的目标，它调节血管内皮生长因子（VEGF）的内皮细胞迁移。这些结论得到了一系列证实性实验的支持。首先，通过二维凝胶电泳和质谱，我们确定膜联蛋白1作为一种蛋白质，其磷酸化是由VEGF诱导的，并通过抑制p38受损。其次，使用体外激酶测定和体内磷酸化测定，我们发现VEGF介导的p38通路下游LIM激酶1的激活触发膜联蛋白1的磷酸化。第三，在小干扰RNA介导的膜联蛋白1敲低后，基质胶中VEGF诱导的细胞迁移和管形成受到抑制。第四，在表达对小干扰RNA敲低不敏感的膜联蛋白1构建体的细胞中，这两个过程都被拯救。最后，VEGF/膜联蛋白1介导的细胞迁移通过抑制p38而受损。因此，我们得出结论，膜联蛋白1的磷酸化调节血管生成的效果，这是与VEGF的p38/LIM激酶1轴的激活。
In this study, we obtained evidence indicating that annexin 1 is a new target of the p38/MAPKAP kinase-2 pathway and that it regulates endothelial cell migration in response to vascular endothelial growth factor (VEGF). These conclusions are supported by a series of substantiating experiments. First, by two-dimensional gel electrophoresis and mass spectrometry, we identified annexin 1 as a protein whose phosphorylation is induced by VEGF and is impaired by inhibiting p38. Second, using in vitro kinase assays and in vivo phosphorylation assays, we found that VEGF-mediated activation of LIM kinase 1 downstream of the p38 pathway triggers the phosphorylation of annexin 1. Third, VEGF-induced cell migration and tube formation in Matrigel are inhibited following small interfering RNA-mediated knockdown of annexin 1. Fourth, both processes are rescued in cells expressing an annexin 1 construct insensitive to the small interfering RNA knockdown. Finally, the VEGF/annexin 1-mediated cell migration is impaired by inhibiting p38. We therefore conclude that phosphorylation of annexin 1 regulates the angiogenic effect that is associated with the activation of the p38/LIM kinase 1 axis by VEGF.