A neurotoxic phospholipase A 2 variant: Isolation and characterization from eastern regional Indian cobra ( Naja naja) venom

Abstract

CM-Sephadex C-25 column chromatography profile of Indian cobra ( Naja naja) venom from eastern region showed a distinct and a dominant phospholipase peak, peak-10, while it was not seen in either southern or western venom samples. Peak-10 was subjected to CM-Sephadex C-25 and Sephadex G-50 column chromatography to isolate NN-X-PLA 2. NN-X-PLA 2 is a single chain protein with the relative molecular weight of 10 kDa by SDS-PAGE. It was toxic to mice with an LD 50 value 0.098 mg/kg body weight (i.p.) and the mice exhibited acute neurotoxic symptoms. Upon indirect stimulation, it inhibited the twitching of frog's gastrocnemius muscle in a dose dependent manner. NN-X-PLA 2 was weakly anticoagulant and devoid of cytotoxicity, myotoxicity, hemorrhage, edema inducing, and directlytic activities and effects on platelet aggregation process. Upon chemical modification independently with p-bromophenacyl bromide and acetic anhydride, NN-X-PLA 2 lost both enzymatic and toxic properties.