Coagulant proteinase from Bothrops colombiensis venom

Area of Science:

• Biochemistry
• Pharmacology
• Toxicology

Background:

• Bothrops colombiensis venom, like other Crotalidae venoms, possesses thrombin-like activity.
• Snake venom components are extensively studied for their biochemical properties and potential therapeutic applications.

Purpose of the Study:

• To purify and characterize the coagulant proteinase from Bothrops colombiensis venom.
• To evaluate the enzyme's specific activity, stability, and potential as a therapeutic agent.

Main Methods:

• Purification using ion-exchange chromatography (DEAE-Sephacel) and gel filtration (Sephadex G-100).
• Characterization using acrylamide gel electrophoresis, flat gel isoelectric focusing, and SDS-PAGE.
• Assay of coagulant activity on bovine fibrinogen and determination of LD50 in mice.

Main Results:

• A purified coagulant proteinase with a specific activity of 231 NIH units/mg (13-fold increase) was obtained.
• The enzyme is a glycoprotein with a native molecular weight >90,000 Da and a monomer molecular weight of ~67,000 Da.
• Optimal coagulation pH is 7.0, with maximal stability between pH 5-6.
• The intraperitoneal LD50 in mice was 4.0 mg/kg.

Conclusions:

• The purified coagulant proteinase from Bothrops colombiensis venom is a potent enzyme with thrombin-like activity.
• Its biochemical properties are comparable to other snake venom coagulant proteinases.
• The enzyme demonstrates potential for development as a therapeutic agent.