Work

Here we describe techniques to purify two proteolitic enzymes out of Echis multisquamatis’s venom. These hydrolases cleave fibrin(ogen) and prothrombin, two key proteins of haemostasis system. Ecamulin, the enzyme that activates prothrombin, was used to obtain an intermediate project of activation – mezothrombin, which in turn activated Factor XIII of blood coagulation, cleaved fibrinopeptides from fibrinogen and induced polymerization of fibrin, and changed platelet aggregation. It can therefore be used to obtain a stable preparation of mezothrombin for in vitro modeling. Ecamulin is unique in its ability to activate both intact prothrombin and its functionally inactive forms (FIFP) and so a promising tool for tests for FIFP levels. We proved that ecamulin test provided reliable data for prognoses of thrombophilia development during systemic lupus erythematosus, stomach ulcer; it is also allows us to control individual sensitiveness of patients to indirect anticoagulants and to choose the adequate dosage. The detection of FIFP is an important marker of protein-synthetics function of liver and allows to determine the rate of liver disfunction and the efficiency of antihepatotoxic therapy. The fibrinogenase from the venom of Echis multisquamatis is highly specific to the ВβR42-A43 peptide bond of fibrinogen. So it was used for the study of molecular structure of fibrinogen and fibrin and conformational re-arrangement during its polymerization of the latter. It was also shown that fibrinogen after truncation by fibrinogenase was unable to normal polymerization, is more sensitive to plasmin digestion, and less potent to support platelet aggregation. Taking into consideration highlighted above properties of fibrinogenase and the fact that fibrinogenase is not act on other haemostasis components, we can conclude that this enzyme could be an effective pro-fibrinolytic, anti-thrombotic agent for the correction of imbalance in haemostasis.