Figure 2.

Effect of Z-VAD-FMK, E64 and leupeptin on cd-LmjMCA enzymatic activity. Protein extracts from Δyca1 yeast cells transfected with the pESC-His vector alone (vector control) and expressing the catalytic domain of LmjMCA (cd-LmjMCA) wild type (wt), and its respective H147A and C202A catalytic site mutants were evaluated for their specific activity with the Ac-VRPR-AMC substrate in the absence or the presence of 100 μM of Z-VAD-FMK, E64 and leupeptin. The AMC release was measured every 15 min for 2 h to determine the activity as the slope of the resulting linear regression. Relative activity was calculated as the fold increase relative to the activity of the vector control (with and without protease inhibitors). Means and S.D. are indicated. ^*P<0.05