Activation of J77A.1 Macrophages by Three Phospholipases A2 Isolated from Bothrops atrox Snake Venom
Figure 1
(a) Chromatographic profile of Bothrops atrox venom. The crude venom of B. atrox after being solubilized was applied on CM-Sepharose (400 mm × 10 mm) previously equilibrated with Tris 50 mM pH 7.4 and eluted under a gradient of NaCl (0–1 M) in the same buffer, in 5 column volumes. The fractions with PLA2 activity are identified with (*). (b) High performance chromatographic profile. Fraction 6 obtained from CM-Sepharose was solubilized in 0.1% TFA (solvent A) and applied on a C18 column (discovery 25 mm × 46 mm, 5 μ, 300 Å) previously equilibrated with the same buffer and eluted with 0.1% TFA in 99.9% acetonitrile (solvent B) under a gradient 0–70% and flow of 1 mL/min. Both chromatograms were monitored with absorbance at 280 nm. (c) SDS-PAGE 12. 5%: samples: 1 molecular weight marker; 2 BthTX-I; 3 fraction 8; 4 fraction 9; 5 fraction 10; 6 11 fraction (BaTX-I); 7 fraction 12. (d) SDS-PAGE 12.5% in reducing conditions: electrophoretic analysis of basic Asp49 PLA2 from B. atrox. Samples: 1 molecular weight; 2 BthTX-I; 3 BaTX-II.
