Skip to main content
. Author manuscript; available in PMC: 2016 Oct 1.
Published in final edited form as: Bioorg Chem. 2015 Jul 13;62:64–73. doi: 10.1016/j.bioorg.2015.07.003

Figure 2. GOAT octanoylation assay optimization.

Figure 2

(A) Reaction mixtures containing 10 μM Ghrelin27, 1 μM octanoyl-CoA, 50 μM palmitoyl-CoA, and 25 μg microsome protein were incubated at 37°C for the indicated time and then quenched in 2% SDS.

(B) Activity over time for 10 μM Ghrelin27 at 25°C and 30°C.

(C) Activity over time for 10 μM Ghrelin27 at 30°C, with 50 μM palmitoyl-CoA in the reaction mixture.

(D) Activity over time for 10 μM Ghrelin27 at 30°C, without 50 μM palmitoyl-CoA in the reaction mixture.

(E,F) Activity in 1 min assay at 30°C with indicated concentration of palmitoyl-CoA. Error bars in F (range of duplicates) are smaller than the data points.

(G) Activity in 1 min at 30°C assay containing 50 μM palmitoyl-CoA with the indicated amount of microsome protein added to the reaction mixture.