UGT1A8 (A–D), UGT1A9 (E–H) and UGT2B7 (I and J) bacculosomes (10 μg) were incubated in the presence of increasing concentrations (1 to 1000 μM) of the racemic (R/S) mixture of bicalutamide (R/S, A, B, E, F and J), or of the pure (R) (R, C, G and I) or (S) enantiomers (S, D and H) and UDPGA (1 mM) for 2 hours at 37°C. The formation of (R)bicalutamide-glucuronide (R-G, A, C, E and G) and (S)bicalutamide-glucuronide (S-G, B, D, F and H) was analyzed by LC-MS/MS. For each panel, large graphs represent the rate of product formation (Y-axis) versus substrate concentration (X-axis) of 2 experiments performed in triplicates (Experiments 1: ·········· and 2: - - - - -) and their mean ( —— ); while small graphs correspond to the mean Eadie-Hofstee plots (rate of product formation (ratio area/min/mg proteins) versus rate of product formation/substrate concentration (ratio area/min/mg proteins/mM).
G: glucuronide.