| | Molecule Name/
Site Name | Km (uM) | kcat (1/s) | Ratio
(k2/k3) | Enzyme Type | Substrate | Product |
| 1 |
Enzyme Activity:
MAPKKthr
Enzyme Molecule:
MAPKK* | 0.0462964 | 0.3 | 4 | explicit E-S complex | MAPK-tyr
| MAPK*
|
| | Rate consts same as for MAPKKtyr. |
| 2 |
Enzyme Activity:
MAPKKtyr
Enzyme Molecule:
MAPKK* | 0.0462964 | 0.3 | 4 | explicit E-S complex | MAPK
| MAPK-tyr
|
| | The actual MAPKK is 2 forms from Seger et al JBC 267:20 14373(1992) Vmax = 150nmol/min/mg From Haystead et al FEBS 306(1):17-22 we get Km=46.6nM for at least one of the phosphs. Putting these together: k3=0.15/sec, scale to get k2=0.6. k1=0.75/46.6nM=2.7e-5 |
| 3 |
Enzyme Activity:
Raf*-GTP-Ras.1
Enzyme Molecule:
Raf*-GTP-Ras | 0.159096 | 0.3 | 4 | explicit E-S complex | MAPKK
| MAPKK-ser
|
| | Kinetics are the same as for the craf-1* activity, ie., k1=1.1e-6, k2=.42, k3 =0.105 These are based on Force et al PNAS USA 91 1270-1274 1994. These parms cannot reach the observed 4X stim of MAPK. So lets increase the affinity, ie, raise k1 10X to 1.1e-5 Lets take it back down to where it was. Back up to 5X: 5.5e-6 |
| 4 |
Enzyme Activity:
Raf*-GTP-Ras.2
Enzyme Molecule:
Raf*-GTP-Ras | 0.159096 | 0.3 | 4 | explicit E-S complex | MAPKK-ser
| MAPKK*
|
| | Same kinetics as other c-raf activated forms. See Force et al PNAS 91 1270-1274 1994. k1 = 1.1e-6, k2 = .42, k3 = 1.05 raise k1 to 5.5e-6 |
| 5 |
Enzyme Activity:
Raf-GTP-Ras.1
Enzyme Molecule:
Raf-GTP-Ras | 0.159096 | 0.3 | 4 | explicit E-S complex | MAPKK
| MAPKK-ser
|
| | Kinetics are the same as for the craf_1* activity, ie., k1=5.5e-6, k2=0.42, k3 = 0.105 These are basedo n Force et al PNAS USA 91 1270-1274, 1994., but k1 is scaled up 5x (ie., Km is scaled down 5x to the value used here and for craf_1* activity: Km = 0.1591). |
| 6 |
Enzyme Activity:
Raf-GTP-Ras.2
Enzyme Molecule:
Raf-GTP-Ras | 0.159096 | 0.3 | 4 | explicit E-S complex | MAPKK-ser
| MAPKK*
|
| | Kinetics are the same as for the craf_1* activity, ie., k1=5.5e-6, k2=0.42, k3 = 0.105 These are basedo n Force et al PNAS USA 91 1270-1274, 1994., but k1 is scaled up 5x (ie., Km is scaled down 5x to the value used here and for craf_1* activity: Km = 0.1591). |
