Biochemical and pharmacological characterization of three toxic phospholipase A2s from Daboia russelii snake venom

Three isoenzymes of phospholipase A(2) (PLA(2)), VRV-PL-VII, and VRV-PL-IX were isolated from Daboia russelii snake venom. The venom, upon gel filtration on Sephadex G-75 column, resolved into six peaks (DRG75 I-VI). The VRV-PL-IIIc was purified by subjecting DRG75II to homogeneity by rechromatography in the presence of 8 M urea on Sephadex G-75 column. The other two isoenzymes VRV-PL-VII and VRV-PL-IX were purified by subjecting DRG75III to ion exchange chromatography on CM-Sephadex C-25 column. Mol wt. for the three PLA(2)s, VRV-PL-IIIc, VRV-PL-VII, and VRV-PL-IX are 13.003 kDa, 13.100 kDa and 12.531 kDa respectively. The VRV-PL-IIIc is not lethal to mice up to 14 mg/kg body weight but it affects blood sinusoids and causes necrosis of the hepatocytes in liver. It causes hemorrhage in kidney and shrinkage of renal corpuscles and renal tubules. The LD(50)s for VRV-PL-VII and VRV-PL-IX are 7 and 7.5 mg/kg body weight respectively. They induced neurotoxic symptoms similar to VRV-PL-V. All the three PIA2s are anticoagulant and induced varying degree of edema in the foot pads of mice. VRV-PL-V and VRV-PL-VII are shown to act as pre and post synaptic toxins, while VRV-PL-IX acts as presynaptic toxin. This is evident from experiments conducted on cultured hippocampal neurons by patch clamp electrophysiology. (C) 2014 Elsevier Inc. All rights reserved.