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. 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 6. GOAT-catalyzed reactions of Ser3 and Dap3 Ghrelin10 substrates at pH 6–9.

Figure 6

(A–C), Reactions were carried out for 1 min in 100 mM Bis-Tris Propane at the indicated pH using Ghrelin10 (Ser3) and Dap3-Ghrelin10. Rates for Ser3 and Dap3 at 7.5 μM are shown in (A) and 100 μM in (B). 100 μM is saturating conditions (see Figure 4C). The ratio of the rate of the two substrates across pH 6–9 is shown in (C). (D) Expression of mouse GOAT in SF9 cells with C-terminal 3xFlag tag and the indicated mutations. 10 μg microsomes (BCA) were loaded in each lane. Ctrl, control microsomes made with empty baculovirus. 3xFlag Std. is purified CK2α-3xFlag. (E) Reactions were carried out for 5 min in 100 mM Bis-Tris Propane at pH 8.0 with 10 μM Ghrelin27 (Ser3) or Dap3-Ghrelin27 with 50 μg total microsome protein.