Table 1. Affinity and fluorescence change for the binding of different nucleoside diphosphates to tetramethylrhodamine-labelled ParM.
Equilibrium dissociation constants (Kd) and fluorescence changes (F+/F−) upon nucleotide binding were determined by fluorescence titration as shown in Figure 1. Data were obtained at 20°C in 30 mM Tris/HCl (pH 7.5), 3 mM MgCl2, 5 μM BSA and 25 mM KCl or 150 mM KCl where indicated.
| 5-ATR-ParM | 6-ATR-ParM | |||
|---|---|---|---|---|
| Nucleotide | F+/F− | Kd (μM) | F+/F− | Kd (μM) |
| GDP | 10.6 | 75.0 | 9.5 | 91.2 |
| GDP (150 mM KCl) | 11.7 | 45.8 | 10.1 | 48.5 |
| ADP* | 14.9 | 30.2 | 14.5 | 37.0 |
| ADP (150 mM KCl) * | 15.2 | 15.3 | 12.3 | 15.8 |
| dGDP | 9.0 | 105 | 8.7 | 149 |
| dADP | 13.1 | 27.5 | 12.2 | 41.7 |
| CDP | 9.5 | 826 | 4.8 | 760 |
| dCDP | 10.6 | 707 | 7.4 | >1000 |
| UDP | 11.6 | 101 | 7.2 | 96.4 |
| TDP | 14.1 | 29.0 | 15.1 | 45.0 |
*Values are from [12].