Table 5.
variant | varied substrate | Km,app (μM) | kcat,app (s−1) | (kcat/Km)app (M−1 s−1) | {[(kcat/Km)app-mutant]/[(kcat/Km)app-wild-type]} × 100 (%) |
---|---|---|---|---|---|
wild-typeb | acetyl-CoA | 29 ± 2 | 32 ± 1 | (1.1 ± 0.1) × 106 | 100 |
P27Ab | 105 ± 4 | 0.77 ± 0.02 | (7.4 ± 0.3) × 103 | 0.67 | |
wild-typec | histamine | 520 ± 54 | 30 ± 1 | (5.8 ± 0.6) × 104 | 100 |
P27Ac | 1800 ± 100 | 0.80 ± 0.02 | 440 ± 30 | 0.76 | |
wild-typec | agmatine | 790 ± 100 | 0.68 ± 0.03 | 850 ± 110 | 100 |
P27Ac | 1300 ± 100 | 0.012 ± 0.001 | 9.7 ± 0.7 | 1.1 |
Kinetic constants are reported as ± the standard error (n = 3).
Reaction conditions: 300 mM Tris-HCl (pH 8.0), 150 μM DTNB, a constant saturating concentration of histamine (>3Km,app), and varying concentrations of acetyl-CoA.
Reaction conditions: 300 mM Tris-HCl (pH 8.0), 150 μM DTNB, a constant saturating concentration of acetyl-CoA (>3Km,app), and varying concentrations of amine substrate.