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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 5. Dap3 ghrelin substrates Form an Octanoyl-Amide.

Figure 5

(A) Hydrazine and hydroxylamine cleavage of octanoylated ghrelin. Reaction mixtures were incubated 1 min at 30°C with 10 μM Ghrelin27 or 100 μM Ghrelin10, quenched in 2% SDS, bound to streptavidin resin, and washed. Capped columns were then incubated overnight with 1 mL of 1M hydrazine or hydroxylamine; 1 M MES buffer was used as a control. Beads and supernatants were then scintillation counted.

(B) Ghrelin10 (Ser3) and Dap3-Ghrelin10 were treated with hydrazine or MES as in (A).