A hemorrhagic
metalloprotease, named
BjussuMP-I, was isolated from Bothrops jararacussu
snake venom by a combination of gel filtration on
Sephacryl S-200 (0.01 M Tris-HCl, pH 7.6
buffer) and
Phenyl Sepharose CL-4B chromatography (0.01 M Tris-HCl plus 4 M NaCl, pH 8.6
buffer, followed by a concentration gradient from 4 to 0 M NaCl at 25 degrees C in the same
buffer).
BjussuMP-I is a 60 kDa
protein with a pI approximately 5.5, which induced
hemorrhage after
intradermal injection in mice, with a minimum hemorrhagic dose of 4.0 microg. The hemorrhagic activity of
BjussuMP-I was totally abolished after incubation with a
chelating agent (
EDTA), corroborating the
metal-dependency of this effect.
BjussuMP-I shows proteolytic activity on
casein and
fibrinogen, although having an activity lower than that of crude B. jararacussu
venom and the
metalloprotease neuwiedase isolated from Bothrops neuwiedi
snake venom. It was recognized by anti-
neuwiedase antibodies, with a reaction of partial immunologic identity.
BjussuMP-I also shows bactericidal activity against Escherichia coli and Staphylococcus aureus. This is the first report on the isolation and characterization of a high molecular weight hemorrhagic
metalloprotease (
BjussuMP-I) from B. jararacussu
venom, which may play a relevant role in local and systemic
bleeding which characterizes Bothrops envenomations.