TABLE 1.
Determination of dissociation constant for membrane binding for various lipid binding domains by the EGFP quenching assay
| Proteins | Lipids | Kd (nM)a (n in parenthesis) | |
| Cuvette Assay | Plate Reader Assay | ||
| PDK1-PH-EGFP | POPC/POPS/dabsylPE/PtdIns(3,4,5)P3 (72:20:5:3) | 35 ± 10 (n = 40 ± 10) | 44 ± 15 (n = 50 ± 20) |
| Akt1-PH-EGFP | POPC/POPS/dabsylPE/PtdIns(3,4,5)P3 (72:20:5:3) | 25 ± 15 (n = 40 ± 12) | 38 ± 10 (n = 50 ± 15) |
| ENTH-EGFP | POPC/POPS/dabsylPE/PtdIns(4,5)P2 (72:20:5:3) | 45 ± 10 (n = 35 ± 15) | 30 ± 10 (n = 40 ± 10) |
| SNX27-PX-EGFP | POPC/POPS/dabsylPE/PtdIns(3)P (72:20:5:3) | 130 ± 20 (n = 40 ± 20) | 110 ± 25 (n = 30 ± 15) |
a
Mean ± SD values from triplicate measurements.