Purification and biochemical characterization of a novel hemorrhagic metalloproteinase from horned viper (Cerastes cerastes) venom.
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.
Boukhalfa-Abib
Hinda
H
Laboratoire de Biologie Cellulaire et Moléculaire, Faculté des Sciences Biologiques, Université des Sciences et de Technologie Houari Boumédienne Bab Ezzouar, Alger, Algeria.
Meksem
Ahmed
A
Laraba-Djebari
Fatima
F
D000818:Animals; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D004492:Edetic Acid / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D006470:Hemorrhage / Q000139:chemically induced; D007525:Isoelectric Focusing; D008297:Male; D008666:Metalloendopeptidases / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism / Q000633:toxicity; D051379:Mice; D008970:Molecular Weight; D010618:Phenanthrolines / Q000494:pharmacology; D011480:Protease Inhibitors / Q000494:pharmacology; D011489:Protein Denaturation; D013696:Temperature; D014757:Viper Venoms / Q000201:enzymology*; D017819:Viperidae; D015032:Zinc / Q000378:metabolism
D010618:Phenanthrolines; D011480:Protease Inhibitors; D014757:Viper Venoms; D004492:Edetic Acid; D008666:Metalloendopeptidases; C069317:hemorrhagic metalloproteinase; D015032:Zinc; C025205:1,10-phenanthroline