Table 1.
Fluorescent Ligand | Receptor | Saturation KD (nM) | Kinetic KD (nM) | kon (min−1 M−1) | koff (min−1) |
---|---|---|---|---|---|
VEGF165a-TMR | NanoLuc-VEGFR2 | 2.03 ± 0.51 | 6.64 ± 4.37 | 1.54 × 107 ± 0.38 × 107 | 0.06 ± 0.02 |
VEGF165b-TMR | NanoLuc-VEGFR2 | 9.53 ± 1.36 | 11.3 ± 3.54 | 7.29 × 106 ± 1.84 × 106 | 0.06 ± 0.01 |
VEGF121a-TMR | NanoLuc-VEGFR2 | 5.54 ± 1.34 | 5.75 ± 0.46 | 8.51 × 106 ± 0.81 × 106 | 0.05 ± 0.00 |
VEGF165a-TMR | NanoLuc-NRP1 | 4.41 ± 1.34 | 4.95 ± 1.25 | 7.11 × 107 ± 2.33 × 107 | 0.26 ± 0.05 |
Equilibrium binding parameters for fluorescent VEGF isoforms derived from saturation and kinetic NanoBRET experiments, showing equilibrium dissociation (KD), association rate (kon), and dissociation rate (koff) constants at NanoLuc-VEGFR2 and NanoLuc-NRP1. Data are expressed as mean ± SEM determined from 5 independent experiments.