Effects of MTS reagents on [3H]NBMPR binding and [3H]2-chloroadenosine uptake by cells transfected with hENT1 cysteine mutants
Data shown are the means ± S.E.M. from at least three independent experiments conducted as described in Figs. 2 to 5.
| Cell Line | MTS Reagent | [3H]NBMPR Binding, Bmax | [3H]2-Chloroadenosine Uptake, Vmax | ||
|---|---|---|---|---|---|
| Control | Treated | Control | Treated | ||
| sites/cell × 105 | pmol · μl−1 · s−1 | ||||
| hENT1-WTa | MMTS | 3.9 ± 0.4 | 5.6 ± 0.4* | 10.8 ± 1.1 | 6.4 ± 1.0* |
| MTSET | 3.6 ± 0.4 | 3.2 ± 0.3* | 9.1 ± 1.6 | 11.3 ± 1.7* | |
| MTSES | 4.1 ± 0.5 | 4.0 ± 0.7 | 8.1 ± 1.2 | 7.6 ± 1.1 | |
| C297S | MMTS | 1.2 ± 0.2 | 1.9 ± 0.3* | 6.8 ± 0.8 | 6.6 ± 0.03 |
| MTSET | 1.7 ± 0.4 | 1.3 ± 0.2 | 5.1 ± 1.6 | 8.1 ± 1.8* | |
| MTSES | 1.0 ± 0.2 | 0.9 ± 0.1 | 6.5 ± 1.7 | 6.2 ± 0.2 | |
| C333S | MMTS | 4.3 ± 0.5 | 7.7 ± 1.5* | 7.8 ± 1.6 | 9.8 ± 2.4 |
| MTSET | 3.5 ± 0.4 | 3.0 ± 0.8 | 5.8 ± 1.2 | 5.7 ± 0.8 | |
| MTSES | 5.0 ± 1.1 | 4.6 ± 1.4 | 6.2 ± 0.2 | 6.8 ± 0.9 | |
| C378S | MMTS | 5.1 ± 0.7 | 7.6 ± 1.2* | 17 ± 2.9 | 9.8 ± 1.3* |
| MTSET | 4.5 ± 0.5 | 4.3 ± 0.8 | 12 ± 1.5 | 12 ± 1.2 | |
| MTSES | 4.5 ± 0.6 | 4.2 ± 0.7 | 3.7 ± 0.6 | 3.2 ± 1.2 | |
| C414S | MMTS | 1.5 ± 0.3 | 3.1 ± 0.4* | 6.7 ± 0.9 | 4.6 ± 0.7* |
| MTSET | 2.0 ± 0.3 | 1.1 ± 0.2* | 6.6 ± 0.7 | 5.8 ± 0.5 | |
| MTSES | 2.4 ± 0.4 | 2.4 ± 0.4 | 5.1 ± 0.2 | 5.7 ± 0.6 | |
| C378S/C414S | MMTS | 6.1 ± 0.3 | 10 ± 0.8* | 25 ± 4.2 | 7.8 ± 1.3* |
| MTSET | 3.2 ± 0.3 | 3.0 ± 0.3 | 24 ± 10 | 29 ± 18 | |
| MTSES | 5.6 ± 2.0 | 5.8 ± 2.0 | 15 ± 3.6 | 13 ± 2.5 | |