RGD-dependent binding of TP508 to integrin alphavbeta3 mediates cell adhesion and induction of nitric oxide.
RGD-dependent binding of TP508 to integrin alphavbeta3 mediates cell adhesion and induction of nitric oxide.
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DOI：
10.1160/th09-07-0447
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发表时间：
2010-07
影响因子：
6.7
通讯作者：
Sheller MR
中科院分区：
文献类型：
--
作者：
Derkach DN;Wadekar SA;Perkins KB;Rousseau E;Dreiza CM;Cheung-Flynn J;Ramos HC;Ugarova TP;Sheller MR
TP508是一种含有23个氨基酸的RGD合成肽，代表人类凝血酶原508至530残基，可减轻缺血再灌注损伤中内皮功能障碍的影响。本研究的目的是研究TP508是否与整合素类跨膜受体家族的成员结合，从而导致一氧化氮的合成。固定化的TP508以剂量和α依赖的方式支持内皮细胞和表达βv RGD3的人胚胎肾细胞的黏附。可溶性TP508还可抑制细胞与固定化纤维蛋白原的黏附。功能阻断抗体和表面等离子共振研究证实了αvβ3的参与。在内皮细胞中，TP508处理可诱导一氧化氮的诱导，而一氧化氮的诱导可被αvβ3特异性、功能阻断的单抗LM609所抑制。最后，TP508对大鼠离体主动脉节段的处理增强了卡巴胆碱诱导的血管松弛。这些结果表明，TP508通过与整合素αvβ3依赖RGD的相互作用而产生潜在的治疗效果。
TP508, a 23-amino acid RGD-containing synthetic peptide representing residues 508 to 530 of human prothrombin, mitigates the effects of endothelial dysfunction in ischaemic reperfusion injury. The objective of this study was to investigate whether TP508 binds to members of the integrin family of transmembrane receptors leading to nitric oxide synthesis. Immobilised TP508 supported adhesion of endothelial cells and αvβ3-expressing human embryonic kidney cells in a dose- and RGD-dependent manner. Soluble TP508 also inhibited cell adhesion to immobilised fibrinogen. The involvement of αvβ3 was verified with function-blocking antibodies and surface plasmon resonance studies. Adhesion of the cells to immobilised TP508 resulted in an induction of phosphorylated FAK and ERK1/2. In endothelial cells, TP508 treatment resulted in an induction of nitric oxide that could be inhibited by LM609, an αvβ3-specific, function-blocking monoclonal antibody. Finally, TP508 treatment of isolated rat aorta segments enhanced carbachol-induced vasorelaxation. These results suggest that TP508 elicits a potentially therapeutic effect through an RGD-dependent interaction with integrin αvβ3.