(A) SDS-PAGE of r-Domain V incubated with various proteases at 37°C for 8 hours (lanes 3 to 7) and 24 hours (lanes 8 to 12). r-Domain V was incubated with UK (lanes 3 and 8), t-PA (lanes 4 and 9), thrombin (lanes 5 and 10), factor Xa (lanes 6 and 11), and sTF/VIIa (lanes 7 and 12) in 0.1 mol/L Tris-HCl (pH 7.5) containing 20 mmol/L NaCl and 0.3 mmol/L CaCl2. Molar ratios of r-Domain V to proteases were 50:1. (B) SDS-PAGE of r-Domain V incubated with plasmin. r-Domain V was incubated with plasmin with a molar ratio of r-Domain V to plasmin was 50:1 at 37 °C. Lanes 3 to 6 are samples taken at 0.25, 0.5, 1, and 2 hours, respectively. For comparison, intact r-Domain V and nicked r-Domain V as prepared in the previous paper14 are shown in lanes 1 and 2 in (A) and (B), respectively. The positions of molecular-weight markers are shown at the left of the gel.
