TABLE 1.
Bmax and affinity for CFT and dopamine to DAT WT and mutants
Experiments performed on transient transfected COS7 cells expressing DAT WT or indicated mutants. The Kd and Bmax values of CFT binding and the Ki for DA displacement were calculated from nonlinear regression analysis of [3H]CFT binding in the presence increasing concentrations of unlabeled ligand (CFT or DA) using 11 consecutive concentrations performed in triplicate. The IC50 values used in the estimation of Kd and Ki values were calculated from means of pIC50 values and the S.E. interval from the pIC50 ± S.E. ND, not detectable.
| DAT construct | CFT Bmax | CFT |
DA |
||
|---|---|---|---|---|---|
| Kd | N | Ki | N | ||
| fmol/105 cells | nm | μm | |||
| WT | 284 ± 31 | 18 (16; 21) | 9 | 0.91 (0.66; 1.26) | 5 |
| R85D | ND | ND | 8 | ND | 4 |
| D476R | 32 ± 3 | 6.7 (5.2; 8.5) | 6 | 12 (8.7; 19) | 4 |
| R85D/D476R | 37 ± 4 | 7.6 (5.1; 11) | 4 | 15 (11; 20) | 4 |