Fig. 2.

Action of long-chain acyl-CoAs in stimulating GOAT activity (A and B) by inhibiting deacylation of [³H]octanoyl CoA (C and D). Membranes from Sf9 cells infected with baculovirus encoding His₁₀-GOAT were prepared as in Fig. 1A. Each 50-μl reaction mixture contained 50 μg of membrane protein, 5 μg of proghrelin-His₈, 1 μM [³H]octanoyl CoA (11 dpm/fmol), and the indicated compound. (A and B) After incubation for 5 min at 37°C, the amount of [³H]octanoyl transferred to proghrelin-His₈ was quantified by nickel chromatography and scintillation counting. Each value represents the average of duplicate assays. (C and D) After incubation at 37°C for the indicated time (C) or 5 min (D), the amount of [³H]octanoate formed in the reaction was quantified by TLC analysis as described in SI Experimental Procedures.