FIGURE 1.
Cleavage of IIWT and IIRR by prothrombinase. Reaction mixtures containing 1.4 μm IIWT (A) or IIRR (B), 30 μm PCPS, 60 μm DAPA, and 30 nm Va in Assay Buffer were initiated by the addition of 0.2 nm Xa. Samples were quenched, analyzed by SDS-PAGE following disulfide bond reduction, stained with Colloidal Blue, and visualized as described under “Experimental Procedures.” Lanes 1–18 correspond to reaction times of 0, 0.33, 0.66, 1.0, 1.33, 1.66, 2, 2.33, 2.66, 3, 4, 6, 8, 12, 20, 30, 45, and 65 min. Identities of the relevant species are indicated in the margin, and two gels, aligned using molecular weight markers, are presented in each panel. The IIaA polypeptide species (Scheme 1) runs off the gel with the dye front in this gel system.