Resumo

RODRIGUEZ, Edith, SANDOVAL, Gustavo A. e YARLEQUE, Armando. Aislamiento y caracterización bioquímica de la bilinearina, un factor activador de protrombina del veneno de la serpiente peruana Bothrops bilineatus (loro machaco). Rev. Soc. Quím. Perú, jan./jun. 2012, vol.78, no.1, p.3-13. ISSN 1810-634X.

A prothrombin activator, called bilinearin, has been detected and isolated from Bothrops bilineatus (loro machaco) snake venom. For its isolation, a combination of two chromatographical steps, CM Sephadex C-50 followed by Sephadex G-100 SF in ammonium acetate buffer 05 M pH 7,0. This enzyme was purified 10,4 times with a yield of 78,3%, and a recovery of 7,5%. Furthermore, it was determined that its molecular weight is 23-kDa, and is formed by 2 polypeptide chains, and 8,4% of attached carbohydrates. Bilinearin is a procoagulant enzyme capable of attacking commercial prothrombin in a direct manner without addition of Ca2+ or phospholipids, producing an active thrombin with clotting activity on human plasma. It was also determined that bilinearin is a metalloprotease because of its strong inhibition by EDTA and EGTA. This enzyme was also inhibited by 2-mercaptoetanol and after exposition to 55ºC, although it was only active until 45ºC. Finally, bilinearin is an immunogenic protein as demonstrated by its reaction to polyvalent bothropic antivenom (INS-Peru) using immunodiffusion and immunoelectrophoresis.

Palavras-chave: Bothrops bilineatus; venom; prothrombin; clotting.