Fig. 1.

Screening for UGT enzymes responsible for the formation of 20-HETE glucuronide using recombinant human UGT isoforms. The glucuronide form of 20-HETE was biosynthesized by incubating 20 μM 20-HETE with the indicated recombinant UGT isoform proteins (15 μg) (BD Biosciences), 1.5 mg/ml alamethicin in 0.5 M Tris-HCl buffer (pH 7.4), and 5 mM UDPGA. The glucuronidated metabolite of 20-HETE was detected by scanning daughter ions of the deprotonated glucuronide at m/z 495.Our results represent one independent data set from two separate experiments. Values represent the mean ± SD of triplicates. UGT1A3, -1A9, and -2B7 metabolized 20-HETE more efficiently than other UGTs. Further details are described in the Experimental Procedures.