Exocyst Complex Component 3-like 2 (EXOC3L2) Associates with the Exocyst Complex and Mediates Directional Migration of Endothelial Cells
Exocyst Complex Component 3-like 2 (EXOC3L2) Associates with the Exocyst Complex and Mediates Directional Migration of Endothelial Cells
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DOI：
10.1074/jbc.m110.212209
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发表时间：
2011-07-08
影响因子：
4.8
通讯作者：
Kreuger, Johan
中科院分区：
文献类型：
--
作者：
Barkefors, Irmeli;Fuchs, Peder Fredlund;Kreuger, Johan
胞吐囊泡是一种蛋白质复合物，可确保胞吐囊泡空间靶向质膜。我们展示了从分化小鼠胚胎干细胞培养物中获得的微阵列数据，这些数据识别了萌芽血管中外囊复合物成分 3 样 2 (exoc3l2) mRNA 的上调。通过不同小鼠组织的 qPCR 分析和小鼠脑切片的免疫荧光分析证实了 exoc3l2 的血管表达。我们检测到原代人内皮细胞中 exoc3l2 mRNA 合成对 VEGFA 的反应上调，并且当细胞在三维 I 型胶原蛋白基质上生长时，这种反应会增强。 Myc 标记的 EXOC3L2 与外囊蛋白 EXOC4 共沉淀，EXOC3L2 的免疫荧光检测显示与 EXOC4 和 EXOC7 的部分亚细胞共定位。最后，我们发现 exoc3l2 沉默抑制 VEGF 受体 2 磷酸化和 VEGFA 引导的培养内皮细胞迁移。
The exocyst is a protein complex that ensures spatial targeting of exocytotic vesicles to the plasma membrane. We present microarray data obtained from differentiating mouse embryonic stem cell cultures that identify an up-regulation of exocyst complex component 3-like 2 (exoc3l2) mRNA in sprouting blood vessels. Vascular expression of exoc3l2 is confirmed by qPCR analysis of different mouse tissues and immunofluorescence analyses of mouse brain sections. We detect an up-regulation of exoc3l2 mRNA synthesis in primary human endothelial cells in response to VEGFA, and this response is enhanced when the cells are grown on a three-dimensional collagen I matrix. Myc-tagged EXOC3L2 co-precipitates with the exocyst protein EXOC4, and immunofluorescence detection of EXOC3L2 shows partial subcellular colocalization with EXOC4 and EXOC7. Finally, we show that exoc3l2 silencing inhibits VEGF receptor 2 phosphorylation and VEGFA-directed migration of cultured endothelial cells.