Research and Development Center, Ezequiel Dias Foundation, 30510-010 Belo Horizonte, Brazil.
Arch Biochem Biophys. 2010 Apr 1;496(1):9-20. doi: 10.1016/j.abb.2010.01.010. Epub 2010 Jan 25.
We report the isolation and structure-function relationship of a 23kDa metalloproteinase named atroxlysin-I from the venom of the Peruvian Bothrops atrox (Jergón). Atroxlysin is a P-I metalloproteinase and contains 204 residues. Its proteolytic activity towards dimethylcasein is enhanced by Ca2+ but inhibited by EDTA, dithiothreitol, excessive Zn2+ and alpha2-macroglobulin. Unlike other structurally homologous P-I metalloproteinases, atroxlysin-I causes hemorrhages. To examine its hemorrhagic activity mechanistically, we studied its function in vitro and in vivo. It cleaved the Ala14-Leu15 and Tyr16-Leu17 bonds in oxidized insulin B-chain and specifically hydrolyzed the alpha-chains of fibrin(ogen) in a dose- and time-dependent manner. Atroxlysin-I cleaved plasma fibronectin and other extracellular matrix proteins (collagens I and IV) and the triple-helical fragment CB3 of collagen IV, but did not degrade laminin-111. Complementarily, the laminin and collagen binding integrins alpha7beta1 and alpha1beta1 were cleaved by atroxlysin. Even without catalytic activity atroxlysin-I inhibited collagen- and ADP-triggered platelet aggregation.
我们从秘鲁响尾蛇(Jergón)的毒液中分离出一种 23kDa 的金属蛋白酶,命名为atroxlysin-I,并研究了其结构-功能关系。Atroxlysin 是一种 P-I 金属蛋白酶,含有 204 个残基。其对二甲基酪蛋白的蛋白水解活性被 Ca2+增强,但被 EDTA、二硫苏糖醇、过量 Zn2+ 和 α2-巨球蛋白抑制。与其他结构同源的 P-I 金属蛋白酶不同,atroxlysin-I 会引起出血。为了从机制上研究其出血活性,我们在体外和体内研究了其功能。它可裂解氧化胰岛素 B 链中的 Ala14-Leu15 和 Tyr16-Leu17 键,并特异性地以剂量和时间依赖的方式水解纤维蛋白原(ogen)的α链。Atroxlysin-I 可切割血浆纤维连接蛋白和其他细胞外基质蛋白(胶原 I 和 IV)以及胶原 IV 的三螺旋片段 CB3,但不降解层粘连蛋白-111。补充的是,atroxlysin 还可切割层粘连蛋白和胶原结合整合素α7β1 和 α1β1。即使没有催化活性,atroxlysin-I 也可抑制胶原和 ADP 触发的血小板聚集。