Table 1.
Steady-State Kinetic Constants for Phosphorylation of PDK1-Tide by His6-PDK1(ΔPH).
| two-substrate a | ADP inhibition b | ||||
|---|---|---|---|---|---|
| parameter | data analyzed | eq | value | Eq | value |
| kcat (s-1) | global | 1 | 0.356 ± 0.010 | 6 | 0.334 ± 0.007 |
| appkcat vs [Tide] | 2 | 0.351 ± 0.015 | 2 | 0.336 ± 0.010 | |
| appkcat vs [ATP] | 3 | 0.346 ± 0.006 | 3 | 0.324 ± 0.014 | |
| KdATP (μM) | global | 1 | 0.57 ± 0.24 | 6 | 0.54 ± 0.15 |
| appKmATP vs [Tide] | 4 | 1.6 ± 1.1 | 4 | 0.804 ± 0.231 | |
| appKmTide vs [ATP] | 5 | 1.0 ± 1.4 | 5 | (1.37) c | |
| αKdATP (μM) | global | 1 | (44.3) c | 6 | (42.6) c |
| appkcat vs [ATP] | 3 | 39.3 ± 2.2 | 3 | 43.4 ± 6.0 | |
| appKmATP vs [Tide] | 4 | 40.8 ± 2.6 | 4 | 42.7 ± 0.9 | |
| appKmTide vs [ATP] | 5 | 25 ± 10 | 5 | 59 ± 32 | |
| KdTide (μM) | global | 1 | 1.1 ± 0.5 | 6 | 1.0 ± 0.3 |
| appKmATP vs [Tide] | 4 | 3.2 ± 2.0 | 4 | (1.52) c | |
| appKmTide vs [ATP] | 5 | 2.1 ± 2.2 | 5 | 1.9 ± 4.6 | |
| αKdTide (μM) | global | 1 | (84.3) c | 6 | (78.2) c |
| appkcat vs [Tide] | 2 | 80.7 ± 9.2 | 2 | 80.2 ± 6.2 | |
| appKmATP vs [Tide] | 4 | 64 ± 13 | 4 | 80.6 ± 5.1 | |
| appKmTide vs [ATP] | 5 | 75.7 ± 7.2 | 5 | 81 ± 13 | |
| α | global | 1 | 77 ± 32 | 6 | 78 ± 21 |
| appKmATP vs [Tide] | 4 | (25.2) c | 4 | (53.1) c | |
| appKmTide vs [ATP] | 5 | (39.3) c | 5 | (42.7) c | |
| KiADP (μM) | global | NA d | NA | 6 | 1.2 ± 0.5 |
| appKiADP vs [Tide] | NA | NA | 7 | 7.5 ± 2.7 | |
| βKiADP (μM) | global | NA | NA | 6 | (58.8) e |
| appKiADP vs [Tide] | NA | NA | 7 | (84) e | |
| appKiADP vs [ATP] | NA | NA | 8 | 19.4 ± 2.8 | |
| β | global | NA | NA | 6 | 49 ± 20 |
| appKiADP vs [Tide] | NA | NA | 7 | 11 ± 4 | |
a
The steady-state kinetic values ± S.E. were determined for the rapid-equilibrium random-order ternary complex mechanism depicted in Fig. 2G.
b
The steady-state kinetic values ± S.E. were determined for the rapid-equilibrium random-order ternary complex mechanism with a dead-end E-Tide-ADP complex depicted in Fig. 4G.
c
This value was calculated from the appropriate relationship given between fitted values of either KdATP, αKdATP, KdTide, αKdTide, or α determined in the indicated method of analysis.
d
Not applicable.
e
This value was calculated from the appropriate relationship given between fitted values of KiADP and β determined in the indicated method of analysis.