Table 1.
Phosphate incorporation and phosphorylation-dependent inhibition of Ets-1 and mutants
| Ets-1 protein
|
Treatmenta
|
mol phosphate/mol Ets-1 (mean ± s.d.)
|
Fold inhibitionb (mean ± s.d.)
|
|---|---|---|---|
| Ets-1 | extract | 5.7 ± 0.9 | 16.6 ± 1.3c |
| Flag–Ets-1 | extract, repurified | N.D. | 62.6 ± 8.7d |
| Flag–Ets-1 | extract + ATP, repurified, ± phosphatase | N.D. | 41.9e |
| Ets-1 | CaMKII | 4.4 ± 0.3 | 55.7 ± 9.5 |
| 4S-A | extract | 0.73 ± 0.2 | 1.3 ± 0.2 |
| 4S-A | CaMKII | 1.4e | 3.0d |
| S251A, S257A | extract | 2.0 ± 0.4 | 3.7 ± 1.6 |
| S282A, S285A | extract | 3.3 ± 0.6 | 5.7 ± 1 |
| L429A | extract | 4.3 ± 0.5 | 2.1 ± 0.4 |
| Flag–L429A | extract + ATP, repurified, ± phosphatase | N.D. | 6.2e |
| L429A | CaMKII | 4.9 ± 0.4 | 5.8 ± 2.0 |
| Y307P | extract | 4.2 ± 2.3 | 7.0 ± 3.0 |
Values represent mean of three experiments except where noted otherwise. (N.D.) Not determined
a
Ets-1 proteins were treated with indicated agent ± ATP except where noted otherwise.
b
Fold inhibition = Kd phosphorylated/Kd mock-treated.
c
Represents a minimum estimate (see Materials and Methods).
d
Mean of two experiments.
e
Value from a single experiment.