Figure 4

PANAT-catalysed hydrolysis of acetyl donors. PANAT at 8 μM (Δ), 4 μM (□) and a control using PBS buffer (×) were incubated with: A) p-nitrophenyl acetate (320 μM) or B) AcCoA (320 μM) at 25°C. The control experiments with PANAT (8 μM) and no acetyl donor (○) are also shown. Reactions were performed in quadruplicate at 25°C and pH 7.4 as described in Methods. The absorbance of p-nitrophenol or 5-thio-2-nitrobenzoic acid (produced by reaction with coenzyme A) is shown (y-axis) as a function of time (x-axis).