Table 1. Binding affinity of NQO1 variants for FAD and dicoumarol determined by ITC.
| NQO1 (Ligand) | Kd1 | Kd2 |
|---|---|---|
| WT (FAD) | ~1 nM | 60 nM |
| p.R139W (FAD) | ~3 nM | 70 nM |
| p.P187S (FAD) | 400 nM | N.app. |
| WT (Dicoumarol) | 122 ± 37 nM | N.app. |
| p.R139W (Dicoumarol) | 56 ± 18 nM | N.app. |
| p.P187S (Dicoumarol) | 11 ± 3 μM | N.app. |
Dissociation constants correspond to fittings to a two-sequential binding sites or to a single-set of independent binding sites (N. app. indicates a single set of sites model). FAD binding constants are the average of two independent titrations (from20) and dicoumarol binding constants are best-fits values ± fitting errors from a single titration (Fig. S1).