Background: Viper snake venoms contain a great variety of toxic proteins. These components mediate their toxicity by either stimulating or inhibiting the haemostatic system of human victims or experimental animals, resulting in common clinical complications of blood clotting or uncontrolled haemorrhage. Therefore it is deemed important to isolate the active component(s) from snake venom with kallikrein-like activity.
Results: A kallikrein-like proteinase of Agkistrodon halys pallas snake venom, designated AHP-Ka, was purified by anion exchange chromatography and affinity chromatography. Physicochemical studies showed that the purified enzyme was a 34 kDa monomeric glycoprotein, the molecular weight of which decreased to 26 kDa after deglycosylation with peptide N-glycosidase F (PNGase F). Sequence studies on the NH(2) -terminal region of the protein indicated that AHP-Ka shared a high degree of sequence homology with other serine proteinases from snake venoms. AHP-Ka showed high catalytic activity and kallikrein-like activity on substrates such as arginine esterase BAEE and chromogenic H-D-Pro-Phe-Arg-pNA·2HCl (S-2302) and was inhibited by protease inhibitor phenylmethylsulfonyl fluoride (PMSF).
Conclusion: The results showed that AHP-Ka isolated from A. halys pallas snake venom and purified by anion exchange chromatography and affinity chromatography is in fact a kallikrein-like enzyme.
Copyright © 2011 Society of Chemical Industry.