Figure 3.

ITC titrations of PEP binding to M₁PYK. (Left) Titration in the absence of Ala is shown. The initial concentration of M₁PYK was 52 μM, and the stock concentration of PEP in the syringe was 10 mM. Titration was performed in 50 mM Tris-HCl buffer (pH 9.0), 500 mM KCl, 10 mM MgCl₂, and 0.1 mM EDTA at 25°C. The first titration point was discarded because of dilution with buffer. (Right) Titration in the presence of Ala is shown. Initial concentrations of M₁PYK and Ala were 39 μM and 10 mM, respectively. The stock concentration of PEP in the syringe was 10 mM.