Figure 4. Structures and synergistic binding with NAD⁺ to PHGDH of the substrate 3-PG and the product 3-PHP and their analogues.

(A) For 3-phosphoglycerate (upper panel) as well as 3-phosphohydroxypyruvate (lower panel) and its analogues,the 2-hydroxypropanoic acid, or 2-oxopropanoic acid respectively, moieties are highlighted in bold. (B) Thermal denaturation of 1 μM PHGDH in the presence of 0.2 mM NAD⁺ or 0.2 mM NADH alone and in combination with 0.2 mM substrate analogues, was measured. Graphs represent mean and standard deviation of three independent experiments with two replicates per experiment. Statistical analysis was performed in GraphPad Prism (two way ANOVA). ^*= p < 0.1, ^*** = p < 0.005, ^**** = p < 0.0005.