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Hydrolysis of ITF5924 by zHDAC6-CD2 as detected by NMR. Evolution of DFMO (black), DFAcH (red), and hydrazide (blue) concentrations and the DFAcH-to-hydrazide molar ratio (purple open circles) over time (0 to 60 h). The kinetic experiment was conducted using 5 μM enzyme and 2 mM DFMO in deuterated assay buffer (25 mM d-Tris-DCl, 0.5 mM d-Tris(2-carboxyethyl)phosphine, pH 8.0). The experiment was carried out in singlicate (N = 1).
