Figure 6

A, Hydrolytic activity of CGP on VE under different pH's. VEs were incubated with 5 μg of CGP (total protein) in the same conditions but different pH's. Lane 1, pH 4.0; Lane 2, pH 5.6; Lane 3, pH 7.0; Lane 4, pH 7.6; Lane 5 pH 8.1; Lane 6, pH 9.0; Lane 7, pH 10.0, Lane 8, VE; Lane 9, FE . The products were analyzed by SDS-PAGE 7.5 %. CGP shows hydrolytic activity at a pH between 5.6 and 8.1. The gel was stained with Coomassie blue. B: Hydrolytic activity of CGP on VE under different protease inhibitors. VEs were incubated with 5 μg total proteins of CGP . Lane 1, calcium free; Lane 2, 5 mM EDTA; Lane 3, 0.7 μg/ml pepstatin; Lane 4, 50 μg/ml TLCK; Lane 5, 50 μg/ml TPCK; Lane 6, VE ; Lane 7, FE ; Lane 8, AE (obtained by treating the VE with CGP) Images are representative of five independent experiments. The products were analyzed in SDS-PAGE 7.5 %. The gel stained with Coomassie blue. Hydrolytic activity of CGP was inhibited under free calcium conditions and 5 mM EDTA.