Analysis of the molecular mechanism in regulation of cell migration by alpha2-plasmin inhibitor

纤溶酶原(PG)缺陷小鼠导致细胞迁移受损，伤口愈合延迟。然而，PG缺陷小鼠的纤维蛋白原缺乏纠正了愈合时间。因此，这些结果表明，纤溶酶(PM)对组织纤维蛋白的降解是细胞迁移的关键。血管抑素(AST)被认为是一种内源性新生血管抑制因子。1998年，AST被证明可以抑制内皮细胞的迁移和管状形成。AST是PG的一个片段，含有赖氨酸结合位点(LBS)，PG和PM通过LBS与纤维蛋白、细胞膜和α2-纤溶酶抑制物(α2-PI)结合。1)α2-PI参与组织纤溶的机制分析：当纤维蛋白原或纤维蛋白以细胞外基质的形式存在时，α2-PI通过X111a因子与之交联。因为α2-PI对选择素有很高的亲和力，所以我们有Ana…进一步裂解α2-PI的糖基，经联氨降解和唾液酸酶处理后，通过Gyocomap分析和阴离子交换HPLC对CQ-PI进行了研究，通过RAMM分析和PNA结合推断了低聚糖的结构。结果表明，α2-PI含有1-3个唾液酸的N-连接低聚糖和1个唾液酸的0-连接低聚糖。此外，本研究还表明α-Pl含有唾液酸基-Lewis x结构。2)细胞迁移研究：含有交联型α-Pl的tibrinogen的细胞外基质可减弱内皮细胞的迁移。此外，α2-PI延缓了内毒素刺激的内皮细胞的迁移。此外，α2-Pl似乎中和了AST对细胞迁移的抑制作用，而a-氨基己酸则不能。我们关于成纤维细胞生长因子与纤维蛋白特异性结合的结果表明，α2-PI在伤口愈合的精细调控中发挥着重要作用。较少

Plasminogen (PG) deficient mice result in impaired cell migration and display delayed wound healing. However, fibrinogen deficiency in PG deficient mice corrects healing time. Consequently, these results suggest that the degradation of tissue-fibrin with plasmin (PM) is critical for cell migration. Angiostatin (AST) has been identified as an endogenous inhibitor of neovascularization. In 1998, AST has been shown to inhibit migration and tube formation of endothelial cells. AST, a fragment of PG, contains lysine-binding site (LBS) through which PG and PM bind to fibrin, cell membrane and alpha2-plasmin inhibitor (alpha2-PI). Thereby, we have analyzed the effect of cL2-PI on the PM-mediated cell migration and the inhibitory effect of AST.1) Analysis of the mechanism of alpha2-PI to participate in tissue fibrinolysis : When fibrinogen or fibrin existed asan extracellular matrix, alpha2-PI was crosslinked to this by factor Xllla. Since alpha2-PI had a high affinity to selectin, we have ana … More lyzed the sugar moiety of alpha2-PI.After degradation with hydrazine and treatment with sialidase, cQ-PI was studied by gyocomap analysis and anion exchange HPLC.Structure of oligosaccharides was deduced by RAMM analysis and the PNA binding. In result, alpha2-PI has N-linked oligosaccharides which has 1-3 sialic acids and 0-linked one containing one sialic acid. In addition, this study has suggested that alpha-Pl contains a sialyl- Lewis x structure.2) Cell migration studies : The extracellular matrix which contained tibrinogen with crosslinked alpha2-PIattenuated endothelial cell migration. In addition, alpha2-PI delayed migration of endothelial cells, which were stimulated with lipopolysaccharide. Furthermore, alpha2-Pl appeared to neutralize the inhibitory effect of AST on cell migration, whereas a-amino caproic acid did not. Our results with the report that fibroblast growth factor specifically binds to fibrin indicate that alpha2-PI plays an important role in a fine regulation of wound healing. Less