Caracterización de la enzima similar a trombina del veneno de Bothrops pictus “jergón de costa”

Translated title of the contribution: Characterization of thrombin like enzyme from Bothrops pictus venom

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Abstract

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.

Translated title of the contributionCharacterization of thrombin like enzyme from Bothrops pictus venom
Original languageSpanish
Pages (from-to)652-658
Number of pages7
JournalRevista Peruana de Medicina de Experimental y Salud Publica
Volume32
Issue number4
DOIs
StatePublished - Oct 2015

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© 2015, Instituto Nacional de Salud. All rights reserved.

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