Purification and characterization of low molecular weight fibrinolytic/hemorrhagic enzymes from snake (Bothrops jararaca) venom

• Published in: Enzyme & protein Vol. 47; no. 3; p. 124
• Main Authors: Maruyama, M, Tanigawa, M, Sugiki, M, Yoshida, E, Mihara, H
• Format: Journal Article
• Language: English
• Published: Switzerland 1993
• Subjects: Animals; Bothrops; Chromatography, Gel - methods; Chromatography, Liquid; Crotalid Venoms; Electrophoresis, Polyacrylamide Gel; Fibrinolysis; Hemorrhage - chemically induced; Isoelectric Focusing; Kinetics; Male; Metalloendopeptidases - isolation & purification; Metalloendopeptidases - metabolism; Metalloendopeptidases - toxicity; Molecular Weight; Rats; Rats, Wistar
• ISSN: 1019-6773
• DOI: 10.1159/000468668

Two low molecular weight fibrinolytic/hemorrhagic enzymes, jararafibrase III and jararafibrase IV, were purified from Bothrops jararaca venom using a fast protein liquid chromatography system. The purified jararafibrase III and jararafibrase IV were single chain proteins with molecular weights of 20,400 +/- 500 and 21,200 +/- 400, respectively, by SDS-PAGE. The isoelectric points of jararafibrase III and jararafibrase IV were 9.4 and 6.9, respectively. The activity of the enzyme was inhibited by 1,10-phenanthroline and EDTA, suggesting that both enzymes were metalloproteinases. The specific fibrinolytic activities of jararafibrase III and jararafibrase IV were 7.5 +/- 0.4 and 6.5 +/- 1.6 units/mg protein, respectively. The enzymes induced local hemorrhage in the skin of rats. The minimal hemorrhagic doses of jararafibrase III and IV were 31.0 and 34.0 micrograms/rat, respectively. The enzymes displayed broad substrate specificities like the previously purified jararafibrases I and II. Jararabrases III and IV degraded type-IV collagen, gelatin, laminin and fibronectin into smaller fragments. The specific activities of jararafibrase III for type-IV collagen and gelatin were 7.6 +/- 0.3 and 43 +/- 11 units/mg protein, respectively. The specific activities of jararafibrase IV for type IV-collagen and gelatin were 16.5 +/- 1.2 and 112 +/- 9 units/mg protein, respectively.