Objectives. To perform a biochemical and molecular characterization of the coagulant principle from Bothrops pictus venom. Materials and methods. We amplified the genetic sequence of this enzyme from cDNA and analyzed the homology of its nucleotide sequence and its deduced protein. This enzyme was also purified for N-terminal sequencing of first 20 amino acids and for coagulation assays using human plasma and human fibrinogen. Furthermore, cleavage pattern on fibrinogen was evaluated using SDS-PAGE and defibrinogenant activity on white mice (18-22 g). Finally, associated carbohydrate content, effect of protease inhibitors and chloride ions on its enzymatic activity were analyzed. Results. The Thrombin-like Enzyme from Bothrops pictus showed homology at primary level of structure with other previously reported TLEs from Viperidae family. Minimum Coagulant Dosis (MCD) on plasma and human fibrinogen were 18 and 6 μg, respectively, and its coagulant potency was 131.1 NHI Thrombin units. This TLE was stable under physiological conditions and chloride ions are not necessary for its activity. Detected associated carbohydrates were hexoses (25.76%), hexosamines (13.12%) and sialic acid (0.76%). Phenyl methyl sulphonyl fluoride (PMSF) and dithiothreitol (DTT) were the main inhibitors of its enzymatic activity, but heparin had no inhibitor effect. Conclusions. The coagulant principle of Bothrops pictus venom is a Thrombin-like enzyme.

Título traducido de la contribuciónCharacterization of thrombin like enzyme from Bothrops pictus venom
Idioma originalEspañol
Páginas (desde-hasta)652-658
Número de páginas7
PublicaciónRevista Peruana de Medicina Experimental y Salud Publica
EstadoPublicada - oct. 2015

Nota bibliográfica

Publisher Copyright:
© 2015, Instituto Nacional de Salud. All rights reserved.

Palabras clave

  • Blood coagulation
  • Bothrops
  • Fibrinogen (source: MeSH NLM)
  • Thrombin


Profundice en los temas de investigación de 'Caracterización de la enzima similar a trombina del veneno de Bothrops pictus “jergón de costa”'. En conjunto forman una huella única.

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