CONTRIBUTION OF PROTEASES IN THE HEMOSTATIC ALTERATIONS INDUCED BY BOTHROPS ALTERNATUS VENOM

GAY, CLAUDIA C.; VAN DE VELDE, A; LEIVA, LAURA; BUSTILLO, SOLEDAD

Buenos Aires

Jornada; XVIII Jornadas Anuales Multidisciplinarias. Sociedad Argentina de Biología,; 2016

Sociedad Argentina de Biología

Bothrops alternatus is a medical important snake in central and northern Argentina. Bothropic envenoming causes proteolysis of tissues, bleeding and coagulation disorders. Coagulating enzymes belong to two proteases families: snake venom serine proteinases (SVSPs) with thrombin-like activity and metalloproteinases (SVMPs) mainly as prothrombin activators; these latest representing more of 50% of B. alternates venom components. In this work, the contribution of metalloproteinases in the hemostatic alterations was studied. The whole venom (10mg/mL) was incubated with an inhibitor of SVMPs, EDTA-Na2 (10mM), the excess of inhibitor was removed by passing the mixture on Sephadex G-25 column (venom without inhibitor was subjected to the same process). Clotting time (CT) was recorded using citrated plasma or fibrinogen (3.5mg/mL) incubated with venom or venom-EDTA-Na2 (venom concentration 270 µg/mL).Results showed that venom-EDTA-Na2 was able to cause a delay of 3.5 times of the CT on citrated plasma compared with venom alone. However, venom-EDTA-Na2 did not alter fibrinogen CT. Besides, venom pre-incubated with specific antibodies anti-baltergin (baltergin is a SVMPs isolated from this venom) was assayed on citrated plasma. In this case, CT showed a delay of 3.1 times compared with crude venom. Results suggest the presence of both proteases families in B. alternatus venom: metalloproteinases acting mainly as pro-coagulant factors. Because of SVMPs are the most important components of this venom, they become relevant targets for the development of new therapeutic agents.