Table 3.
Binding parameters of PaDHPase to 5-AU.
| PaDHPase | λ max (nm) | λ em shift (nm) | Quenching (%) | K d value (μM) |
|---|---|---|---|---|
| PaDHPase | From 336.5 to 343 | 6.5 | 87.3 | 97.7 ± 2.0 |
| PaDHPase-Y155A | From 336.5 to 342 | 6.0 | 68.6 | 247.1 ± 4.0 |
| PaDHPase-S289A | From 337 to 339 | 2.0 | 61.6 | 256.0 ± 3.2 |
| PaDHPase-N337A | From 336 to 339.5 | 3.5 | 72.4 | 192.1 ± 8.0 |
| PaDHPase-C318A | From 336 to 342 | 6.0 | 86.8 | 103.3 ± 3.1 |
| Znα-PaDHPase | From 335 to 336.5 | 1.5 | 49.3 | 281.5 ± 9.0 |
The decrease in the intrinsic fluorescence of ScDHOase was measured with a spectrofluorimeter (Hitachi F-2700; Hitachi High-Technologies, Japan). The Kd was obtained using the equation: ΔF = ΔFmax − Kd(ΔF/[5-AU]).