Table 3.
Spectral binding parameter estimates for (S)-warfarin and flurbiprofen.
| (S)-warfarin | Flurbiprofen | |||||
|---|---|---|---|---|---|---|
| KsaμM | ΔAmaxa ×10−3 | ΔA/KbμM−1 × 10−3 | KsaμM | ΔAmaxa ×10−3 | ΔA/KbμM−1 × 10−3 | |
| Wild-type | 8.9 ± 0.8 | 5.3 ± 0.1 | 0.6 (100) | 3.4 ± 0.4 | 18 ± 0.5 | 5.3 (100) |
| F100L | 4.2 ± 0.2 | 6.5 ± 0.06 | 1.5 (262) | 2.2 ± 0.2 | 16 ± 0.3 | 7.3 (137) |
| F114L | 12.2 ± 0.3 | 10 ± 0.07 | 0.8 (139) | 4.1 ± 0.6 | 10 ± 0.4 | 2.4 (46) |
| F476L | 16.3 ± 1.0 | 11 ± 0.2 | 0.7 (114) | 9.1 ± 1.0 | 11 ± 0.4 | 1.2 (23) |
| F114W | 3.2 ± 0.3 | 12 ± 0.3 | 3.8 (636) | 4.0 ± 0.2 | 34 ± 0.4 | 8.5 (160) |
| F476W | 7.0 ± 0.4 | 9.3 ± 0.1 | 1.3 (225) | 18.3 ± 1.9 | 4.7 ± 0.1c | 0.3 (5) |
| F114L/F476L | 34.1 ± 2.4 | 15 ± 0.3 | 0.4 (75) | 9.9 ± 1.3 | 4.4 ± 0.2 | 0.4 (8) |
| F114W/F476W | 6.7 ± 0.5 | 12 ± 0.2 | 1.8 (303) | 22.3 ± 4.1 | 7.3 ± 0.4 | 0.3 (6) |
a
Ks and ΔAmax refer to the dissociation constant and the maximum observed difference (390–420 nm), respectively, as determined by fitting to a one site saturation ligand binding model
b
Bold number in parentheses refers to percent of wild-type ΔA/K
c
Low concentrations demonstrated unusual binding properties, therefore Ks was determined from 5–100 μM, which showed typical type I binding spectra