TABLE 3

Kinetics of NSAID glucuronidation by pooled HLMs

Rates were determined at NSAID concentrations from 10 μM to 1 mM under the following conditions: 1 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. Rates of glucuronidation were fit to the Michaelis-Menten equation and parameters are reported as best-fit value (standard error).


NSAID

Apparent Km

Apparent Vmax

Vmax/Km

Fit
μM pmol/min/mg μl/min/mg R 2
protein protein
Sulindac 75.4 (26.5) 382.9 (35.4) 5.1 0.90
Sulindac sulfone 34.1 (9.1) 663.4 (41.9) 19.5 0.93
Indomethacin 30.0 (6.7) 376.6 (17.3) 12.6 0.96
Ibuprofen 188.3 (16.8) 657.3 (18.5) 3.5 0.99
Flurbiprofen 56.3 (9.7) 446.1 (19.0) 7.9 0.97
Ketoprofen 140.7 (5.7) 742.2 (8.9) 5.3 1.0
Diclofenac 71.6 (23.6) 2685 (150.9) 37.5 0.96
Naproxen
75.3 (15.8)
1708 (93.9)
22.7
0.96