Laboratoire des Venins et Toxines, Institut Pasteur de Tunis, Tunis, Tunisia.
Lab Invest. 2010 Apr;90(4):510-9. doi: 10.1038/labinvest.2009.137. Epub 2010 Feb 8.
Integrins are essential in the complex multistep process of angiogenesis and are thus attractive targets for the development of antiangiogenic therapies. Integrins are antagonized by disintegrins and C-type lectin-like proteins, two protein families from snake venom. Here, we report that CC-PLA2-1 and CC-PLA2-2, two novel secreted phospholipases A(2) (PLA(2)) isolated from Cerastes cerastes venom, also showed anti-integrin activity. Indeed, both PLA(2)s efficiently inhibited human brain microvascular endothelial cell adhesion and migration to fibrinogen and fibronectin in a dose-dependent manner. Interestingly, we show that this anti-adhesive effect was mediated by alpha5beta1 and alphav-containing integrins. CC-PLA2s also impaired in vitro human brain microvascular endothelial cell tubulogenesis on Matrigel and showed antiangiogenic activity in vivo in chicken chorioallantoic membrane assay. The complete PLA(2) cDNAs were cloned from a venom gland cDNA library. Mature CC-PLA2-1 and CC-PLA2-2 contain 121 and 120 amino acids, respectively, including 14 cysteines each and showed 83% identity. Tertiary model structures of CC-PLA2-1 and CC-PLA2-2 were generated by homology modeling. This is thus the first study describing an antiangiogenic effect for snake venom PLA(2)s and reporting first clues to their mechanism of action on endothelial cells.
整合素在血管生成的复杂多步骤过程中是必不可少的,因此成为抗血管生成治疗药物开发的有吸引力的靶标。整合素被蛇毒中的两种蛋白家族——解体素和 C 型凝集素样蛋白拮抗。在这里,我们报告说,从 Cerastes cerastes 毒液中分离得到的两种新型分泌型磷脂酶 A2(PLA2)CC-PLA2-1 和 CC-PLA2-2 也具有抗整合素活性。事实上,这两种 PLA2 都能以剂量依赖的方式有效地抑制人脑血管内皮细胞对纤维蛋白原和纤维连接蛋白的黏附和迁移。有趣的是,我们表明这种抗黏附作用是由 alpha5beta1 和 alphav 整合素介导的。CC-PLA2s 还损害了体外人脑血管内皮细胞在 Matrigel 上的小管形成,并在鸡胚绒毛尿囊膜试验中显示出体内抗血管生成活性。完整的 PLA2 cDNA 是从毒腺 cDNA 文库中克隆出来的。成熟的 CC-PLA2-1 和 CC-PLA2-2 分别含有 121 和 120 个氨基酸,每个都含有 14 个半胱氨酸,并且具有 83%的同源性。通过同源建模生成了 CC-PLA2-1 和 CC-PLA2-2 的三级结构模型。因此,这是首次描述蛇毒 PLA2s 的抗血管生成作用,并首次报道其在血管内皮细胞上的作用机制的线索。