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Membrane binding properties of the PDK1 PH domain WT and mutants
Equilibrium SPR experiments were performed in 20 mm Tris-HCl, pH 7.4, containing 0.16 m KCl, and Kd values were determined as shown in Fig. 1E.
| PDK1 PH domains |
Kd |
||
|---|---|---|---|
| POPC/PtdIns(3,4,5)P3 (97:3) | POPC/POPS (80:20) | POPC/POPS/PtdIns(3,4,5)P3 (77:20:3) | |
| nm | |||
| WT | 52 ± 4 | 95 ± 17 | 30 ± 5 |
| K465A | 1700 ± 160 | 160 ± 50 | 150 ± 25 |
| R466A | 58 ± 9 | 390 ± 180 | 65 ± 20 |
| K467A | 59 ± 8 | 200 ± 25 | 65 ± 10 |
| R466A/K467A | 50 ± 15 | 560 ± 200 | 220 ± 20 |
