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. 2015 Nov 1;290(51):30648–30657. doi: 10.1074/jbc.M115.688523

FIGURE 3.

FIGURE 3.

Activity of Hat1 and Hat1-containing complexes as a function of Hif1 and Asf1 histone chaperones. Enzyme kinetic analysis comparing the activity of Hat1 (red), HAT-B (yellow), and NuB4 (green) on H3-H4 substrate (top and middle panels) and Asf1/H3-H4 substrate (bottom panels). A, the data for Hat1 or Hat1-containing complexes with varying concentrations of H3-H4 (1–75 μm) are fit to Michaelis-Menten kinetics (GraphPad Prism 5) to calculate steady state parameters. B, the data for Hat1 or Hat1-containing complexes varying amounts of H3-H4 (1–20 μm) are fit to the linear portion of the rate data (GraphPad Prism 5) to calculate kcat/Km. C, the data for Hat1 or Hat1-containing complexes with varying concentrations of Asf1/H3-H4 (0.5–2.5 μm)) are fit to the linear portion of the rate data (GraphPad Prism 5) to calculate kcat/Km. All reactions were performed using 100 nm enzyme at saturating levels of 14C-labeled acetyl CoA (500 μm), and all measurements were carried out in duplicate.