Purification and Biochemical Characterization of Three Myotoxins from Bothrops mattogrossensis Snake Venom with Toxicity against Leishmania and Tumor Cells
Chromatographic and electrophoretic profile of Bothrops mattogrossensis venom fractioning. (a) CM-Sepharose columns equilibrated with solvent A (50 mM ammonium bicarbonate, pH 8.0) and eluted with a 0–100% concentration gradient of solvent B (500 mM ammonium bicarbonate, pH 8.0) at a 5.0 mL/minute flow. Emphasis on peak 9, rechromatographed. (b) Rechromatography of fraction 9 on Discovery C18 column equilibrated with solvent A (0.1% TFA) and eluted with a concentration gradient of 0–100% of solvent B (99.9% acetonitrile and 0.1% trifluoroacetic acid) and a 1.0 mL/min flow. Emphasized in blue are fractions 13, 14, and 15 characterized as phospholipases BmatTX-I, BmatTX-II, and BmatTX-III, respectively. Controls: MW: molecular weight standard; BTx: BthTX-I a basic enzymatically inactive PLA2 (Lys49) (10 μg) isolated from Bothrops jararacussu venom. Absorbances read at 280 nm. Electrophoresis gel made with 12.5% (w/w) acrylamide/bis-acrylamide.