title: Purification and characterization of a potent hemolytic toxin with phospholipase A2 activity from the venom of Indian Russell's viper.
abstract: A potent toxin with phospholipase A2 (PLA2) and hemolytic activity in vitro was purified from the Russell's viper venom of eastern India (RVV-EI). The purified protein (RVV-PFIIc') of 15.3 kDa molecular weight, and a lethal toxicity dose (LD50i.p.) of 0.1 mg/kg body weight, was the most toxic PLA2 so far reported from the Indian subcontinent. The material also possessed anticoagulant activity as it enhanced the prothrombin induced plasma clotting time in vitro. The PLA2 toxin (RVV-PFIIc') was shown to be different from other PLA2s of RVV in respect to one or more of these parameters e.g. molecular weight, isoelectric pH, in vivo toxicity, specific activity of the enzyme and certain other biological activities. The first 19 amino terminal sequence (NLFQFAEMIVKMTGKEAVH) of RVV-PFIIc' showed variable degree of homology (42.1-94.7%) with those of other RVV-PLA2s described in the literature. Antisera raised against RVV-EI or RVV-PFIIc', though completely neutralized the in vivo lethal toxicity of RVV-EI or RVV-PFIIc', failed to inhibit their PLA2 activity in vitro thereby suggesting that in vivo toxicity and in vitro activity of the enzyme may not be directly related. Apart from RVV-PFIIc', at least two other PLA2 isozymes were found to be present in RVV-EI that were distinct from RVV-PFIIc' in respect to their molecular, biological as well as serological properties. The significance of these and related data in antivenom therapy is discussed.
authors.affiliation: Department of Microbiology, Bose Institute, Calcutta, India.
authors.forename: Amit K
authors.initials: AK
authors.lastname: Chakraborty
authors.forename: Robert H
authors.initials: RH
authors.lastname: Hall
authors.forename: Asoke C
authors.initials: AC
authors.lastname: Ghose
mesh_terms: D000818:Animals; D000997:Antivenins / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D006461:Hemolysis; D015151:Immunoblotting; D010455:Peptides / Q000737:chemistry; D010741:Phospholipases A / Q000737:chemistry* / Q000302:isolation & purification*; D054467:Phospholipases A2; D017434:Protein Structure, Tertiary; D017840:Daboia; D014757:Viper Venoms / Q000737:chemistry* / Q000201:enzymology
chemical_list: D000997:Antivenins; D010455:Peptides; D014757:Viper Venoms; D010741:Phospholipases A; D054467:Phospholipases A2