Table 2.
FRET analysis of BFP-labeled EGFR-ICD proteins
| Sample | % Energy transfer | Distance (Å) | Kd (μM) |
| EGFR-ICD | 46 ± 1.7 | 40 | 0.21 ± 0.04 |
| EGFR-ICD~Pa | 28 ± 2.3 | 46 | 0.27 ± 0.08 |
| EGFR-Δ976 | 57 ± 1.2 | 37 | 0.16 ± 0.01 |
| EGFR-Δ976~Pa | 58 ± 1.4 | 37 | 0.18 ± 0.01 |
| EGFR-Δ1022 | 43 ± 1.0 | 41 | 0.21 ± 0.01 |
| EGFR-Δ1022~Pa | 41 ± 1.3 | 41 | 0.24 ± 0.01 |
Donor emission spectra of various EGFR-ICD-BFP constructs recorded during TNP-ATP titrations (e.g. Fig. 3 ▶) were integrated to yield relative quantum yields, which were corrected for inner filter quenching effects (Fig. 4 ▶). The quantum yield versus TNP-ATP concentration profiles were fit with hyperbolic curves (Fig. 6 ▶), yielding the percentage of energy transfer at saturation, the corresponding distance in Å ngstroms, and the apparent TNP-ATP dissociation constant (Kd). Percentages of energy transfer and Kd values are given as the average ± SE of three independent determinations.
aDenotes the BFP-labeled proteins under phosphorylating conditions.