FIGURE 4.

Representative radiochromatograms of the total [1-¹⁴C]lipid components of following FAAH reactions in the presence (or absence) of 100 μm PDP-EA or cardanol-EA. Reactions were initiated by the addition of purified rat or At-FAAH protein (0.3 μg) with 50 mm BisTris propane-HCl (pH 9.0) and 100 μm [1-¹⁴C]NAE 12:0 in a final volume of 0.15 ml. Reactions proceeded at 30 °C with shaking (120 rpm) for 25 min. A–C, reaction with At-FAAH protein, solvent control (DMSO), with 100 μm PDP-EA, and with 100 μm cardanol-EA; D–F, reaction with rat FAAH protein, solvent control (DMSO), with 100 μm PDP-EA, and with 100 μm cardanol-EA; G, assays with heat-denatured At-FAAH protein (5 min at 100 °C) plus cardanol-EA (100 μm). Lipids were extracted and separated by TLC (60:40:5; v/v/v). Chromatograms were obtained by radiometric scanning of the TLC plate.