Table 3.
Estimates of kinetic constants for CBL autoubiquitination with UbcH5B
| kcat (min−1) | Km (μM) |
kcat/Km (min−1 M−1) |
|
|---|---|---|---|
| Full-length CBL | |||
| WT1,2 | – | – | 3,400 |
| M222E1,2 | – | – | 8,300 |
| M222F1,2 | – | – | 3,200 |
| WT+ZAP-70 peptide1,2 | – | – | 3,600 |
| CBL47–435 | |||
| WT1,2 | – | – | 190 |
| Y371F1,2 | – | – | 200 |
| Y368F1,2 | – | – | 387 |
| WT1,3 | – | – | 68 |
| pY3713 | 0.68 ± 0.07 | 6.5 ± 1.6 | 105,540 |
| Y368F pY3713 | 0.75 ± 0.07 | 7.7 ± 1.4 | 97,260 |
| Y368F pY371 K389A3 | 0.22 ± 0.04 | 17.4 ± 4.9 | 12,780 |
| Y368F pY371 V431A3 | 0.13 ± 0.02 | 15.6 ± 3.5 | 8,460 |
Kinetic constants were determined by fitting two independent datasets onto the Michaelis-Menten equation. s.e.m. are indicated.
1
An estimated kcat/Km was reported, because the reaction was not saturated.
2
Reactions were performed at 30°C.
3
Reactions were performed at 22°C.