Purification and characterization of low molecular weight fibrinolytic/hemorrhagic enzymes from snake (Bothrops jararaca) venom.
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.
Maruyama
M
M
Department of Physiology, Miyazaki Medical College, Japan.
Tanigawa
M
M
Sugiki
M
M
Yoshida
E
E
Mihara
H
H
D000818:Animals; D017837:Bothrops; D002850:Chromatography, Gel / Q000379:methods; D002853:Chromatography, Liquid; D003435:Crotalid Venoms; D004591:Electrophoresis, Polyacrylamide Gel; D005342:Fibrinolysis; D006470:Hemorrhage / Q000139:chemically induced*; D007525:Isoelectric Focusing; D007700:Kinetics; D008297:Male; D008666:Metalloendopeptidases / Q000302:isolation & purification* / Q000378:metabolism / Q000633:toxicity; D008970:Molecular Weight; D051381:Rats; D017208:Rats, Wistar
D003435:Crotalid Venoms; D008666:Metalloendopeptidases; C089473:jararafibrase III; C089474:jararafibrase IV