Kinetics of NSAID glucuronidation by selected UGT Supersomes
NSAID | UGT | Apparent Km | Apparent Vmax | Vmax/Km | Fit |
|---|---|---|---|---|---|
| μM | pmol/min/mg protein | μl/min/mg protein | R 2 | ||
| Sulindac | 1A1a | 5.2 (1.3) | 31.7 (1.1) | 6.1 | 0.92 |
| 1A3a | 19.6 (3.6) | 99.2 (3.4) | 5.1 | 0.97 | |
| 1A9a | 68.0 (8.7) | 8.0 (0.3) | 0.1 | 0.98 | |
| 2B7b | 124.4 (25.6) | 229.2 (17.8) | 1.8 | 0.96 | |
| Sulindac sulfone | 1A1b | 5.2 (1.4) | 459.6 (18.9) | 88.3 | 0.95 |
| 1A3a | 15.0 (4.7) | 429.8 (23.6) | 28.7 | 0.91 | |
| 1A9a | 44.6 (10.3) | 7.5 (0.4) | 0.2 | 0.93 | |
| 2B7b | 34.1 (9.1) | 1211 (76.5) | 35.6 | 0.98 | |
| Indomethacin | 1A1c | 13.2 (2.8) | 118.3 (6.3) | 9.0 | 0.98 |
| 1A3c | 3.1 (0.7) | 10.2 (0.4) | 3.3 | 0.97 | |
| 1A9c | 68.5 (16.5) | 127.1 (15.0) | 1.9 | 0.99 | |
| 2B7c | 17.7 (4.7) | 229.4 (16.9) | 13.0 | 0.97 | |
| Ibuprofen | 1A3a | 413.2 (28.7) | 146.5 (4.0) | 0.4 | 1.0 |
| 1A9a | 98.4 (19.6) | 219.4 (12.9) | 2.2 | 0.96 | |
| 2B4a | 171.0 (30.2) | 57.5 (3.1) | 0.3 | 0.98 | |
| 2B7a | 30.0 (6.9) | 266.5 (12.8) | 8.9 | 0.95 | |
| Flurbiprofen | 1A3b | 201.8 (40.2) | 472.9 (39.6) | 2.3 | 0.98 |
| 1A9b | 42.7 (6.4) | 263.1 (11.3) | 6.1 | 0.98 | |
| 2B4a | 86.4 (16.0) | 213.2 (10.6) | 2.5 | 0.96 | |
| 2B7a | 6.1 (1.2) | 217.6 (5.9) | 35.7 | 0.96 | |
| Ketoprofen | 1A3a | 136.5 (17.6) | 688.7 (25.9) | 5.0 | 0.99 |
| 1A9a | 76.4 (15.3) | 163.3 (8.6) | 2.1 | 0.95 | |
| 2B4a | 126.3 (10.7) | 404.2 (9.9) | 3.2 | 0.99 | |
| 2B7a | 11.7 (2.5) | 789.9 (30.1) | 67.5 | 0.96 | |
| Diclofenac | 1A3c | 16.7 (6.2) | 715.6 (73.0) | 42.9 | 0.95 |
| 1A9c | 7.7 (1.8) | 375.7 (19.9) | 48.8 | 0.97 | |
| 2B4c | 4.7 (0.9) | 370.5 (13.0) | 78.8 | 0.98 | |
| 2B7c | 19.9 (4.1) | 1323 (79.8) | 66.7 | 0.98 | |
| Naproxen | 1A3a | 322.5 (67.6) | 778.9 (60.0) | 2.4 | 0.98 |
| 1A9a | 96.0 (20.9) | 909.0 (54.3) | 9.5 | 0.95 | |
| 2B4a | 195.3 (32.3) | 960.6 (50.3) | 4.9 | 0.98 | |
|
| 2B7b | 7.0 (1.9) | 1613 (75.7) | 230.4 | 0.95 |
↵ a Rates fit Michaelis-Menten equation from 1 to 1000 μM concentrations of NSAID.
↵ b Rates fit Michaelis-Menten equation from 1 to 500 μM concentrations of NSAID.
↵ c Rates fit Michaelis-Menten equation from 1 to 100 μM concentrations of NSAID. Rates were determined at NSAID concentrations from 1 μM to 1 mM under the following conditions: 0.5 mg/ml protein, 50 μg/mg alamethicin, 1 mM PMSF, 8.5 mM saccharolactone, 2 mM UDP-glucuronic acid, and 10 mM MgCl2 in 50 mM potassium phosphate, pH 7. Kinetic parameters are reported as best-fit value (standard error).