TABLE 3.
Norepinephrine (NE) uptake and Nisoxetine (NX) binding affinity at WT and mutant hNET.
| Position | [3H]-NE Km (μM) | [3H]-NX Kd (nM) | [3H]-NE Vmax (%) | [3H]-NX Bmax (%) | (Vmax/Bmax) (%) | |
| WT | 1.580.33 | 3.670.54 | 100 | 100 | 100 | |
| F72A | TMH1a | 2.501.29 | 6.353.15 | 14.835.36* | 16.493.48* | 90.08.84 |
| A73G | Loop1a-1b | 7.741.59* | 7.584.38 | 41.203.66* | 95.337.44 | 43.011.10* |
| A73L | Loop1a-1b | 0.631.97 | 5.280.59 | 4.670.69* | 102.606.80 | 4.607.49* |
| A73R | Loop1a-1b | 1.090.29 | 5.311.99 | 0.650.47* | 21.174.93* | 3.105.40* |
| A73V | Loop1a-1b | 4.800.58* | 4.391.09 | 58.304.06* | 164.216.43* | 35.510.49* |
| A77L | TMH1b | 5.101.10* | 2.380.91 | 1.020.58* | 145.305.47* | 0.706.05* |
| A77R | TMH1b | 1.500.30 | 22.475.62* | 7.302.47* | 88.275.00 | 8.007.47* |
| N78A | TMH1b | 0.930.69 | 7.173.92 | 17.051.05* | 96.501.50 | 18.02.55* |
| V148A | TMH3 | 2.741.28 | 26.338.78* | 28.795.95* | 82.800.61 | 35.06.56* |
| N153A | TMH3 | 3.650.09* | 14.215.43 | 3.621.82* | 94.0013.34 | 3.8515.16* |
| I156A | TMH3 | 1.920.77 | 5.753.87 | 5.050.83* | 11.240.76* | 45.01.59* |
| G320A | TMH6 | 3.120.13* | N.D. | 8.700.50* | 6.681.83* | 130.22.33* |
| V325A | TMH6 | 1.380.44 | N.D. | 88.735.76 | N.D. | N.D. |
| F329A | TMH6 | 0.900.39 | 16.504.50 | 4.600.65* | 8.595.24* | 53.05.89* |
| N350A | TMH8 | 0.360.22* | 45.165.22* | 3.301.40* | 112.5012.50 | 2.9013.90* |
| S420A | TMH8 | 0.450.04* | 7.922.06 | 65.8711.45* | 149.2522.45* | 44.133.90* |
| G423A | TMH8 | 5.960.89* | 11.931.41 | 78.531.35 | 120.408.24 | 65.29.59* |
| M424A | TMH8 | 0.830.09* | 7.054.85 | 68.507.65 | 83.704.09 | 81.811.74* |
N.D., not determined. Values are the mean ± SEM, of 3–4 independent experiments for hNET mutants and n = 15 for WT, each performed in triplicate. *Significant mutant effects compared with WT hNET.