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. Author manuscript; available in PMC: 2018 Nov 15.
Published in final edited form as: J Am Chem Soc. 2017 Nov 1;139(45):16222–16227. doi: 10.1021/jacs.7b07603

Figure 1.

Figure 1

Bpa incorporation in HDAC8 does not dramatically affect hydrolytic activity. (A) Positions for Bpa incorporation were chosen based on proximity to a coumarin-linked peptide substrate (Ac-Arg-His-Lys(ac)-Lys(ac)-methylcoumarin) visualized in the co-crystal structure of HDAC8 (PDB ID: 2V5W). (B) Measurement of the activity of HDAC8-Bpa mutants assayed using Fluor-de-Lys assay with a coumarin-linked peptide substrate. Assays were performed using 50µM peptide and 1µM Co2+-HDAC8. The value of kcat/Km (app) was calculated assuming a linear dependence on peptide concentration.