Purification and characterization of an antiplatelet peptide, arietin, from Bitis arietans venom.

By means of Fractogel TSK-50, CM-Sephadex C-50 column chromatography, gel filtrations on Sephadex G-75 and Sephacryl S-200 columns and reverse-phase HPLC, an antiplatelet peptide, arietin, was purified from venom of Bitis arietans. Arietin was shown to be an Arg-Gly-Asp-containing peptide with a NH2-terminus, Ser-Pro-Pro-Val-Cys-Gly-Asn-Lys- (Mr 8500). Arietin dose-dependently inhibited aggregation of human platelet suspension stimulated by ADP, thrombin, collagen and U46619 with IC50, 1.3-2.7.10(-7) M, while it had no effect on the initial shape changes and only slightly affected ATP release of platelets caused by thrombin and collagen. Arietin also blocked platelet aggregation in platelet-rich plasma and whole blood, and inhibited thrombin-induced clot retraction of platelet-rich plasma. Furthermore, arietin (6.5.10(-8) M) completely blocked the fibrinogen-induced aggregation of elastase-treated platelets, indicating that arietin interferes with the fibrinogen binding to fibrinogen receptors on platelet membranes. In conclusion, arietin, an Arg-Gly-Asp-containing peptide, inhibits platelet aggregation probably through the blockade of fibrinogen binding to the activated platelets.