Purification and biochemical characterization of a novel hemorrhagic metalloproteinase from horned viper (Cerastes cerastes) venom.

Hinda Boukhalfa-Abib, Ahmed Meksem, Fatima Laraba-Djebari

Laboratoire de Biologie Cellulaire et Moléculaire, Faculté des Sciences Biologiques, Université des Sciences et de Technologie Houari Boumédienne Bab Ezzouar, Alger, Algeria.

Comparative biochemistry and physiology. Toxicology & pharmacology : CBP 2009 Aug

filter terms:

Snake venoms contain metalloproteinases that contribute to the local effects observed after envenoming. In this study, a hemorrhagic metalloproteinase (CcH1) was purified from Cerastes cerastes venom by a combination of gel filtration, ion exchange, affinity and RP-HPLC chromatography. The hemorrhagin was homogeneous on SDS-PAGE, with a molecular mass of 25 kDa. Isoelectric focusing revealed a pI of 5.5. CcH1 displayed hemorrhagic and proteolytic activities, but no esterolytic activity. The hemorrhagic and proteolytic activities of CcH1 were inhibited by EDTA and 1,10-phenanthroline, but not by PMSF, suggesting that this protein is a zinc-metalloproteinase. Furthermore, the hemorrhagic and proteolytic activities of CcH1 were stable in solution at up to 40 degrees C, with a loss of activity at > or =70 degrees C. The molecular mass and the inhibition assays suggest that the metalloproteinase CcH1 belongs to class P-I of SVMPs.

Citation

Hinda Boukhalfa-Abib, Ahmed Meksem, Fatima Laraba-Djebari. Purification and biochemical characterization of a novel hemorrhagic metalloproteinase from horned viper (Cerastes cerastes) venom. Comparative biochemistry and physiology. Toxicology & pharmacology : CBP. 2009 Aug;150(2):285-90