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. 2005 Jan 23;11(2):160–166. doi: 10.1038/nm1179

Figure 2. Differences in GPIase and peptidase activities of ACE.

Figure 2

(a) Effects of metal chelating reagents on the wild-type Ace. EDTA, CyDTA or EGTA was applied to the PLAP conversion assay and the released PLAP was detected by immunoblotting (top). Bottom panels indicate effects of these reagents on the peptidase activity (activity under control condition; i.e., no reagent, was considered 1.0). Dose effect of EDTA on GPIase and peptidase activities. EDTA was applied at 0.1 mM or 1 mM (left). Effects of high dose (1 mM) CyDTA and EGTA on GPIase and peptidase activities (right). Buffer indicates no reagent applied. (b) GPIase and peptidase activities of the wild-type (WT), E414D and H413K-H417K mutants. GPIase activity is expressed as released PLAP activity (left). The dipeptidyl carboxypeptidase (Dipeptidase) activity of the same samples (right). ND, not detected. (c) Effects of metal chelating reagents on the H413K-H417K mutant. EDTA, CyDTA or EGTA was applied to the PLAP conversion assay and the released PLAP was detected by immunoblotting. Dose effect of EDTA on GPIase activity (left). EDTA was applied at 0.1 mM or 1 mM. Effects of high-dose (1 mM) CyDTA and EGTA on GPIase activity (right).