Artículos de revistas
Purification from Bothrops lanceolatus (fer de lance) venom of a fibrino(geno)lytic enzyme with esterolytic activity
Registro en:
Toxicon. Pergamon-elsevier Science Ltd, v. 36, n. 5, n. 745, n. 758, 1998.
0041-0101
WOS:000073963500006
10.1016/S0041-0101(97)00118-9
Autor
De Araujo, AL
Donato, JL
Bon, C
Institución
Resumen
Bothrops lanceolatus venom has high caseinolytic, phospholipasic, esterolytic and hemorrhagic activities. In spite of having no coagulant effect on plasma, this venom contains a thrombin-like enzyme. Using gel filtration and ion-exchange chromatographies, we have purified an esterolytic fraction (F-II-la) from this venom with a protein yield of 4% and a 58% recovery in enzyme activity. SDS-PAGE in the presence of beta-mercaptoethanol showed that the enzyme is a single chain polypeptide with a M-w = 38,100. Immunodiffusion and immunoelectrophoresis of fraction F-II-la against serum from horses immunized with B. lanceolatus venom and against rabbit antiserum prepared using fraction F-II-la both showed a single immunoprecipitin line. The K-m and V-max values for TAME hydrolysis were 0.85 mM and 38.6 mu mol/min/mg, respectively. The esterolytic activity was completely inhibited by PMSF (10 mM) but not by EDTA (20 mM). Fraction F-II-la hydrolyzed the alpha and beta chains of fibrinogen. Degradation of the a chain occurred within 10 min while that of the beta-chain was slower. The enzyme had no effect on the gamma-chain even after 4 h of hydrolysis. (C) 1998 Elsevier Science Ltd. All rights reserved. 36 5 745 758