Isolation of anticoagulant from the venom of tick, Boophilus calcaratus, from Uzbekistan

Boophilus calcaratus is a tick found in Central Asia and a common parasite to domestic animals. Venom from this tick was fractionated by two-step column chromatography, Sephadex G-75, and DEAE-Sephadex A-25. The homogeneity of the anticoagulant was examined by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The purified component is named calcaratin and has a molecular weight of 14,500. The effect of the purified anticoagulant component (calcaratin) on various sites of the blood coagulation cascade scheme was examined and compared with crude venom. The chromogenic substrates S-2238 (H-D-Phe-Pip-Arg-pNA 2HCl) for thrombin and S-2765 (N-alpha-Z-D-Arg-Gly-Arg-pNA 2HCl) for factor Xa were also investigated. Activated partial clotting times were all prolonged, suggesting the anticoagulation nature of the purified component and crude venom. Prolongation of fibrinogen clotting time (FCT) is highly suggestive of the antithrombin property of the purified component and its original venom.