Fig 4. Hydrophobic chromatographic fractioning of Tityus serrulatus MMG homogenate.

MMG homogenate was fractioned with 50% ammonium sulfate on a HiTrap Butyl column (GE) equilibrated in 50 mM phosphate buffer (pH 6.0). Elution was performed using a gradient of 1.7–0 M ammonium sulfate in the same buffer. (A) Activity of each fraction against 10 μM Z-FR-MCA was measured in 100 mM Tris-HCl buffer (pH 8.0) containing 10 mM CaCl₂ (●) or in the presence of 5.0 mM benzamidine (○). (B) The activity of each fraction against 10 μM Z-FR-MCA was measured in 100 mM CP-buffer (pH 5.5) containing 3.0 mM cysteine and 3.0 mM EDTA in the absence (●) and presence of different peptidase inhibitors: (□) 10 μM E-64; (○) 1.0 mM PMSF; (Δ) 10 μM pepstatin.