Laboratory of Endothelial Cell Biology, Institut de recherches cliniques de Montréal (IRCM), Université de Montréal, Montreal, Quebec, Canada.
Mol Cell. 2010 Aug 13;39(3):468-76. doi: 10.1016/j.molcel.2010.07.013.
Disruption of adherens junctions between endothelial cells results in compromised endothelial barrier function and in altered angiogenesis. Nitric oxide (NO) produced by endothelial NO synthase (eNOS) is essential for increased vascular permeability induced by vascular endothelial growth factor (VEGF). However, the molecular mechanisms by which NO modulates endothelial permeability remain elusive. Here, we show that, within adherens junctions, beta-catenin is a substrate for S-nitrosylation by NO. Stimulation of endothelial cells with VEGF induces S-nitrosylation of beta-catenin, which is dependent on expression and activity of eNOS. Furthermore, VEGF-induced S-nitrosylation of beta-catenin is inhibited in eNOS(-/-) mice. We identify Cys619, located within the VE-cadherin interaction site, as the major S-nitrosylation locus in response to VEGF. Inhibition of S-nitrosylation at Cys619 prevents NO-dependent dissociation of beta-catenin from VE-cadherin and disassembly of adherens junction complexes and inhibits VEGF-stimulated endothelial permeability. Thus, we identify S-nitrosylation of beta-catenin as a modulator of intercellular contacts between endothelial cells.
细胞间黏附连接的破坏会导致内皮细胞屏障功能受损，并改变血管生成。内皮型一氧化氮合酶（eNOS）产生的一氧化氮（NO）对于血管内皮生长因子（VEGF）诱导的血管通透性增加是必不可少的。然而，NO 调节内皮通透性的分子机制仍不清楚。在这里，我们表明，在黏附连接中，β-连环蛋白是 NO 介导的 S-亚硝基化的底物。VEGF 刺激内皮细胞可诱导β-连环蛋白的 S-亚硝基化，这依赖于 eNOS 的表达和活性。此外，eNOS(-/-)小鼠中 VEGF 诱导的β-连环蛋白 S-亚硝基化被抑制。我们确定了位于 VE-钙粘蛋白相互作用位点内的 Cys619 作为 VEGF 反应的主要 S-亚硝基化位点。Cys619 的 S-亚硝基化抑制阻止了 NO 依赖性β-连环蛋白从 VE-钙粘蛋白解离以及黏附连接复合物的解体，并抑制了 VEGF 刺激的内皮通透性。因此，我们确定β-连环蛋白的 S-亚硝基化是内皮细胞细胞间连接的调节剂。
