Purification and Characterization of a Phospholipase A 2 and Identification of a kappa Bungarotoxin from Bungarus sindanus sindanus (Sindhi Krait) Snake Venom (original) (raw)
A Phospholipase A 2 termed as BSS PLA 2 was purified from the venom of Bungarus sindanus sindanus (Sindhi Krait) through two steps: size exclusion chromatography on sephadex G-75 and reversed-phase chromatography on Vydac C18 HPLC column. The homogeneity and molecular mass of BSS PLA 2 were analysed by SDS-PAGE and MALDI-TOF mass spectrometry. The N-terminal sequence of 32 amino acids of the BSS PLA 2 revealed close resemblance with other species of bungarus snakes. The BSS PLA 2 showed an optimum activity in the alkaline range (pH 8.0-9.5), while the optimum temperature stability was found to be 60 °C. The effect of calcium ion (Ca 2+) concentration revealed that in the absence of Ca 2+ the enzyme showed some activity however, the enzymatic activity increases with increase in Ca 2+ concentration. The addition of exogenous metal ions, including Mg 2+ with Ca 2+ increases, while that of Ba 2+ with Ca 2+ slightly decreases the enzymatic activity of BSS PLA 2. Additionally, kappa bungarotoxin with the mass of 7358.5 Da was identified by MALDI TOF MS/MS from another peak of the same HPLC elution. Two of the tryptic peptides matched with kappa-6-bungarotoxin when searched through Mascot search engine.
Loading Preview
Sorry, preview is currently unavailable. You can download the paper by clicking the button above.