Table 1.
Substrates and the reaction conditions for each enzymatic assay.
| Substrates | (mM) | pHa | DTT (mM) | EDTA (mM) | CaCl2 (mM) | NaCl (mM) | MnCl2 (mM) | |
|---|---|---|---|---|---|---|---|---|
| PEP (EC 3.4.21.26) | Z-Gly-Pro-β-NA | 0,125 | 7,4 | 2 | — | — | — | — |
| PGI (EC 3.4.19.3) | pGlu-β-NA | 0,125 | 7,4 | 2 | 2 | — | — | — |
| APBb (EC 3.4.11.6) | Arg-β-NA | 0,125 | 6,5 | — | — | — | 150 | — |
| APN/CD13b (EC 3.4.11.2) | Ala-β-NA | 0,125 | 7,4 | 1,2 | — | — | — | — |
| AlaAP (EC 3.4.11.14) | Ala-β-NA | 0,125 | 7,4 | 1,2 | — | — | — | — |
|
| ||||||||
| APA (EC 3.4.11.7) | H-Glu-β-NA | 0,125 | 7,4 | 0,65 | — | 0,5 | — | — |
| AspAP (EC 3.4.11.21) | Asp-β-NA | 0,125 | 7,4 | — | — | — | — | 1 |
| DPPIV/CD26 (EC 3.4.14.5) | H-Gly-Pro-β-NA | 0,2 | 8,3 | — | — | — | — | — |
| CAP (EC 3.4.11.3) | L-Cys-di-β-NA | 0,125 | 5,9 | — | — | — | — | — |
| NEPc (EC 3.4.24.11) | N-Dansyl-D-Ala-Gly-pNO2-Phe-Gly | 0,125 | 7,4 | — | — | — | — | — |
aReaction buffers: Phosphate buffer (50 mM) was used for PEP, PGI, APB, APN/CD13, and AlaAP. Tris-HCl (50 mM) was used for APA, AspAP, DPPIV/CD26, CAP, and NEP. b40 μM of the AlaAP inhibitor puromycin was added; c4.6 nM of the angiotensin-converting enzyme inhibitor captopril was added.