Dissociation constant of MD380
| Dissociation constant | ||||
| Construct | No nucleotide | AMP-PNP | ADP-Vi | ADP |
| μM | μM | μM | μM | |
| WT | 1.1 ± 0.1 | 1.0 ± 0.1 | 2.8 ± 0.6 | 0.8 ± 0.1 |
| α-R403A | 4.2 ± 0.4 | 4.2 ± 0.2 | 4.7 ± 0.5 | 2.5 ± 0.2 |
| α-E416A | 4.3 ± 0.3 | 3.8 ± 0.3 | 4.0 ± 0.3 | 2.8 ± 0.2 |
| Dissociation constant | ||||
| Construct | No nucleotide | AMP-PNP | ADP-Vi | ADP |
| μM | μM | μM | μM | |
| WT | 1.1 ± 0.1 | 1.0 ± 0.1 | 2.8 ± 0.6 | 0.8 ± 0.1 |
| α-R403A | 4.2 ± 0.4 | 4.2 ± 0.2 | 4.7 ± 0.5 | 2.5 ± 0.2 |
| α-E416A | 4.3 ± 0.3 | 3.8 ± 0.3 | 4.0 ± 0.3 | 2.8 ± 0.2 |
Errors indicate the errors in curve fitting. In wild-type MTs, the maximum binding Bmax was calculated to be 0.76, 0.76, 0.77, and 0.74 for no nucleotide, AMP-PNP, ADP-Vi, and ADP conditions, respectively. These values were used as the Bmax for mutants measured in the same nucleotide conditions.