Table 1.
RFK and FMN residual activities of CaFADS in the presence of the HTS hits.
| % Residual RFK activity |
|||||
|---|---|---|---|---|---|
| ≤ 5 | 5–50 | 50–75 | ≥75 | ||
| % Residual FMNAT activity | |||||
| ≤5 | 9, 29, 33, 37 | 14,47 | 2, 11, 15, 17, 19, 24, 27, 31, 43 | ||
| 5–50 | 35 | 38 | 22,32 | 1, 3, 4, 5, 6, 7, 8, 10, 12, 13, 16, 18, 25, 28, 39, 40, 44, 46 | |
| ≥50 | – | – | – | – | |
Values measured at 25 °C, in 20 mM PIPES, pH 7.0, 2.5% DMSO and 0.8 or 10 mM MgCl2 when assaying the RFK or the FMNAT activities, respectively. The final concentration of each HTS hit was 250 μM and saturating concentrations of all the substrates were used. The compounds highlighted in italics completely inhibited the FMNAT activity with minor effects on the RFK one, and were selected as FMNAT hits.