KD values for MAP, PEITC, and coumarin binding to CYP2A6, CYP2A13, and mutants of CYP2A13 in which individual active site residues have been substituted with the residue found at the corresponding position in CYP2A6
Numbers in italics give the -fold difference from the CYP2A13 value for the corresponding ligand. Values represent the mean of two titrations for CYP2A6, CYP2A13, and all mutants with ≥3-fold difference from the parental enzyme.
Enzyme | MAP | PEITC | Coumarin | ||||||
|---|---|---|---|---|---|---|---|---|---|
| KD | KD/KD (2A13 wt.) | KD | KD/KD (2A13 wt.) | KD | KD/KD (2A13 wt.) | ||||
| μM | μM | μM | |||||||
| CYP2A6 | 74 | 74 | 6.1 | 14 | 3.1 | 1.1 | |||
| CYP2A13 | 1.0 | 1.0 | 0.43 | 1.0 | 2.7 | 1.0 | |||
| CYP2A13 mutants | |||||||||
| L110V | 1.6 | 1.6 | 0.46 | 1.1 | 1.7 | 0.63 | |||
| A117V | 4.1 | 4.1 | 1.2 | 2.8 | 1.8 | 0.67 | |||
| S208I | 12.0 | 12. | 1.4 | 3.3 | 7.9 | 2.9 | |||
| A213S | 2.9 | 2.9 | 0.87 | 2.0 | 3.4 | 1.3 | |||
| F300I | 15.0 | 15 | 6.0 | 14. | 14.0 | 5.2 | |||
| A301G | 8.2 | 8.2 | 3.1 | 7.2 | 4.2 | 1.6 | |||
| M365V | 1.7 | 1.7 | 1.7 | 4.0 | 5.0 | 1.9 | |||
| L366I | 0.56 | 0.56 | 0.34 | 0.79 | 0.57 | 0.21 | |||
| G369S | 1.6 | 1.9 | 1.8 | 4.2 | 6.0 | 2.2 | |||
| H372R | 0.77 | 0.77 | 1.0 | 2.3 | 1.5 | 0.56 | |||
| CYP2A6 mutants | |||||||||
| I208S/I300F/G301 | 12.0 | 12. | 4.9 | 11 | 6.0 | 2.2 | |||
| I300F/G301A/S369G | 12.3 | 12 | 1.9 | 4.4 | 3.1 | 1.1 | |||
| I208S/I300F/G301A/S369G | 3.5 |
3.5
| 2.1 |
4.8
| 2.3 |
0.85
| |||