Soluble perlecan domain i enhances vascular endothelial growth factor-165 activity and receptor phosphorylation in human bone marrow endothelial cells
Soluble perlecan domain i enhances vascular endothelial growth factor-165 activity and receptor phosphorylation in human bone marrow endothelial cells
复制标题
DOI：
10.1186/1471-2091-11-43
复制
发表时间：
2010-11-03
期刊：
影响因子：
--
通讯作者：
Gomes, Ronald R., Jr.
中科院分区：
文献类型：
--
作者：
Muthusamy, Arivalagan;Cooper, Carlton R.;Gomes, Ronald R., Jr.
背景：固定化重组串珠素结构域I（PlnDI）在体外结合并调节肝素结合生长因子的活性。然而，尚未报告PlnDI溶液中的活动。在这项研究中，我们评估了可溶形式的能力，以调节血管内皮生长因子-165（VEGF（165））增强毛细血管样形成，和VEGF受体-2磷酸化的人骨髓内皮cells，在vitro.Results：在溶液中，PlnDI结合VEGF（165）在硫酸乙酰肝素和pH依赖性的方式。外源性PlnDI可增强毛细血管样形成;然而，PlnDI/VEGF（165）混合物联合收割机可增强形成，超过单独PlnDI或VEGF（165）刺激的形成。PlnDI还刺激VEGF受体-2磷酸化，并且PlnDI/VEGF的混合物（165）减少峰值VEGF受体-2磷酸化（Tyr-951）所需的时间，并增加Akt磷酸化。PlnDI结合固定的神经纤毛蛋白-1和VEGF受体-2，但对神经纤毛蛋白-1具有更大的亲和力。PlnDI与神经纤毛蛋白-1结合，但不与VEGF受体-2结合依赖于修饰PlnDI的硫酸乙酰肝素链。有趣的是，VEGF（165）而不是VEGF（121）的存在显著增强PlnDI与Neuropilin-1和VEGF受体-2的结合。结论：我们的观察表明可溶形式的PlnDI具有生物活性。此外，PlnDI硫酸乙酰肝素链单独或与VEGF（165）一起可以在体外增强VEGFR-2信号传导和血管生成事件。我们建议在基底膜或细胞外基质周转过程中释放的PlnDI可能具有类似的活性，在体内。
Background: Immobilized recombinant perlecan domain I (PlnDI) binds and modulates the activity of heparin-binding growth factors, in vitro. However, activities for PlnDI, in solution, have not been reported. In this study, we assessed the ability of soluble forms to modulate vascular endothelial growth factor-165 (VEGF(165)) enhanced capillary tube-like formation, and VEGF receptor-2 phosphorylation of human bone marrow endothelial cells, in vitro.Results: In solution, PlnDI binds VEGF(165) in a heparan sulfate and pH dependent manner. Capillary tube-like formation is enhanced by exogenous PlnDI; however, PlnDI/VEGF(165) mixtures combine to enhance formation beyond that stimulated by either PlnDI or VEGF(165) alone. PlnDI also stimulates VEGF receptor-2 phosphorylation, and mixtures of PlnDI/VEGF(165) reduce the time required for peak VEGF receptor-2 phosphorylation (Tyr-951), and increase Akt phosphorylation. PlnDI binds both immobilized neuropilin-1 and VEGF receptor-2, but has a greater affinity for neuropilin-1. PlnDI binding to neuropilin-1, but not to VEGF receptor-2 is dependent upon the heparan sulfate chains adorning PlnDI. Interestingly, the presence of VEGF(165) but not VEGF(121) significantly enhances PlnDI binding to Neuropilin-1 and VEGF receptor-2.Conclusions: Our observations suggest soluble forms of PlnDI are biologically active. Moreover, PlnDI heparan sulfate chains alone or together with VEGF(165) can enhance VEGFR-2 signaling and angiogenic events, in vitro. We propose PlnDI liberated during basement membrane or extracellular matrix turnover may have similar activities, in vivo.