Purification, Characterization And Anti-thrombus Effect Of A Plasminogen Activator From Gloydius Brevicaudus Venom

In 1993,A novel plasminogen activator form Trimeresurus stejnegeri venom(TSV—PA) has been identified and purified by Yun Zhang for the first time.It offers the new subject for developing new medicine from venom..This study was designed to isolate and purify novel plasminogen activator from Gloydius brevicaudus venom(GBV)and analyse its characterization and biological activities.1. Purification and characterization of the plasminogen activator from Gloydius brevicaudus Venom.1.1 Purification of GBV-PAAffinity chromatography of Gloydius brevicaudus Venom in Benzamidine Sepharose 6B resulted in the separation of two protein peaks. The fractionⅡcan specifically activate plasminogen through an enzymatic reaction estimated by the fibrin plate method. GBV-PA was purified from the fractionⅡby Lichrospher C18 4.6/250 reverse chromatography.1.2 The characterization of GBV-PAIt was homogeneous as a single band in both reducing and non-reducing showed on SDS-polyacrylmide gel electrophoresis (SDS-PAGE,Tris system) with molecular weight 40KDa as calculated by Image Master VDS system.The isoelectric point of GBV-PA was pH 5.2 by disc polyacrylamide gel eletrophoresis.2. The biological activities of GBV-PA2.1 The specific activity of GBV-PAGBV-PA specifically activates plasminogen through an enzymatic reaction that leads to plasmin generation and displays an indirect fibrinolysis activity through using the chromogenic substrate method. The plasminogen activating activity was 2.87 t-PA IU/mg according the chromogenic substrate S- 2390.