Table 1.
Masked antagonists show protease-dependent binding and activity
| SPR | Cell assay | |||
|---|---|---|---|---|
| kon (M−1 s−1)/103 | koff (s−1) × 105 | KD (nM) | IC50 (nM) | |
| 4 | 96.3 (n.r.) | 189 (n.r.) | 19.7 (n.r.) | 149 ± 17.8* |
| 4 + TEV | 522 (n.r.) | 181 (n.r.) | 3.5 (n.r.) | 64 ± 10.7* |
| 5 | 30 ± 16.7 | 264 ± 116 | 122 ± 91 | 1285 ± 214 |
| 5 + TEV | 364 ± 10.4 | 159 ± 7.6 | 4.4 ± 0.3 | 582 ± 171 |
| 7 | 33.1 ± 15.4 | 155 ± 8.5 | 61.4 ± 43.3 | 566 ± 342 |
| 7 + TEV | 408 ± 78.7 | 321 ± 6 | 8.1 ± 1.8 | 34 ± 26.7 |
| 9 | 66.6 ± 5.3 | 145 ± 11 | 21.8 ± 0.5 | 212 ± 10.4* |
| 9 + TEV | 441 ± 67.9 | 311 ± 9.6 | 7.2 ± 1 | 24 ± 4.8* |
| 10 | 10.4 ± 2.5 | 238 ± 82.6 | 243 ± 113 | 1143 ± 102 |
| 10 + TEV | 392 ± 117 | 410 ± 26.7 | 11.1 ± 3.2 | 25 ± 8.5 |
| 15 | 9.3 ± 2.5 | 181 ± 50.1 | 205 ± 68.3 | 743 ± 68.7 |
| 15 + TEV | 425 ± 67.7 | 355 ± 8.2 | 8.5 ± 1.2 | 30 ± 12.8 |
| 16 | 209 (n.r.) | 122 (n.r.) | 5.8 (n.r.) | N/A |
| 16 + TEV | 459 (n.r.) | 199 (n.r.) | 4.3 (n.r.) | N/A |
| 19 | 10.3 ± 3.3 | 211 ± 52.8 | 228 ± 102 | 1606 ± 45.3 |
| 19 + TEV | 478 ± 32.3 | 283 ± 4.4 | 5.9 ± 0.5 | 54 ± 32.9 |
| 20 | 9.6 ± 1.5 | 109 ± 20.3 | 114 ± 13.8 | 909 ± 33 |
| 20 + TEV | 290 ± 65.8 | 332 ± 7.8 | 11.7 ± 2.4 | 31 ± 15.7 |
| No | 1,165 ± 617 | 262 ± 113 | 2.5 ± 1 | 16 ± 6.6 |
| No + TEV | 400 ± 18.2 | 363 ± 29.3 | 9.1 ± 0.9 | N/A |
Table of mean kinetic parameters for the masked antagonists before and after protease treatment including NoMA (No) for the SPR binding kinetic assay (Left) and the cellular activity assay (Right). For the binding kinetics, error represents SD whereas some samples were only tested once (no replicate, n.r.). For the cell assay, error represents either the SD in the average IC50 across multiple experiments (SI Appendix, Fig. S6) or the 95% CI of three replicates within a single experiment (*).