Table 1.
NAMPT protein |
Km (μM) |
Vmax (nmol/min/mg) |
Vmax/Km |
Ki (μM) |
||
---|---|---|---|---|---|---|
Nama | PRPPb | PRPP | Nam | PRPP | ||
Wildtype | <0.1 | 0.46 ± 0.13 | 123 ± 2 | 586 | 103 ± 22 | 52 ± 13 |
α-Loop double mutant K(68/71)S | <0.1 | 0.28 ± 0.08 | 122 ± 14 | 381 | 93 ± 19 | 93 ± 24 |
α-Loop triple mutant K(68>73)A | <0.1 | 0.29 ± 0.05 | 108 ± 23 | 512 | 69 ± 19 | 104 ± 20 |
β-Loop double mutant K(42/48)A | <0.1 | 0.73 ± 0.14 | 148 ± 13 | 487 | 166 ± 41 | 103 ± 23 |
β-Loop full mutant K/R(42>51)A | <0.1 | 0.89 ± 0.14 | 110 ± 11 | 177 | 183 ± 52 | 905 ± 225 |
β-Loop deleted mutant ΔK(42>51)c | <0.1 | 0.12 ± 0.01 | 0.6 ± 0.02 | 5 | ND | ND |
172 | 780 ± 40 | 1.3 ± 0.05 | 0.0017 | ND | ND |
Abbreviation: ND, not detectable.
At 0.1 mM PRPP, 1 mM ATP.
At 5 μM Nam, 1 mM ATP.
Upper row shows kinetic parameters for the first phase (i.e., Km1, Vmax1), lower row for the second stage of kinetics (Km2, Vmax2).