The fluorescence intensity (FI) at varying temperatures (upper panel) and derivative melt curves calculated by differences in FI at each temperature (lower panel) are shown. Peak temperature in the lower panel of graph dFl/dT is considered as Tm. The recombinant proteins are (A) GST, (B) ERM-BP-WT, (C) D12A, (D) ERM-BP-DBM, (E) K12A and (F) C198A. Different concentrations of NAD+ were used as indicated as 0 mM (blue), 1 mM (red), 2 mM (green) and 4 mM (pink). Experiments were done in quadruplicate for each NAD+ concentration for each protein and in three independent experiments. Representative thermal shift assay is shown from one experiment. (G) Thermal stability assay of ERM-BP-WT with different amounts of NADP as indicated 0 mM (green), 1 mM (blue), 2 mM (pink) and 4 mM (orange) in quadruplicate for each NADP concentration.