TABLE 2.
Ki values of DOG for binding to the single and quadruple mutant RasGRP2 C1 domain and to the full-length quadruple mutant RasGRP2 as well as to the RasGRP1 C1 domain and the full-length RasGRP1
The single and quadruple C1 domains were fused with GST and expressed in a bacterial system, the quadruple full-length RasGRP2 was fused to HaloTag and expressed in HEK293 cells, and the full-length RasGRP1 was fused with MBP and expressed in bacteria. After isolation of the proteins, the binding of DOG was determined in a competitive binding assay with [3H]PDBu. Values represent the mean ± S.E. (n = 3 independent experiments).
| Receptor | Ki |
|---|---|
| nm | |
| hRasGRP2 C1 | |
| S8Y | 5240 ± 680 |
| N7T/S8Y/A19G/I21L | 60.7 ± 7.7 |
| hRasGRP1 C1 | 51 ± 14 |
| hRasGRP2 full length | |
| N7T/S8Y/A19G/I21L | 414 ± 88 |
| hRasGRP1 full length | 107.3 ± 9.3 |