Table 1.
Affinity and kinetic interaction parameters determined by SPR
| MET (1051–1349) protein variants | Tepotinib |
|||
|---|---|---|---|---|
| KDa (nM) | kdb (s−1) | kac (M−1 s−1) | Residence timed (min) | |
| WT (no ATP preincubation) | NA | <0.0001 | NA | >160 |
| WT (after ATP preincubation) | 5.7 | 0.0095 | 1,670,000 | 1.8 |
| WT mix (species C1) | 0.08 | 0.00012 | 1,490,000 | 138 |
| WT mix (species C2) | 2.4 | 0.0064 | 2,690,000 | 2.6 |
| Y1234E/Y1235E | 2.9 | 0.0079 | 2,755,000 | 2.1 |
| Y1235D | 2.3 | 0.0057 | 2,500,000 | 2.9 |
| F1200I (species G1) | 0.5 | 0.0057 | 11,500,000 | 2.9 |
| F1200I (species G2) | 23.4 | 0.0208 | 888,150 | 0.8 |
| L1195V (species H1) | 0.3 | 0.0009 | 3,060,000 | 18.5 |
| L1195V (species H2) | 80.3 | 0.0345 | 430,000 | 0.5 |
| Y1230C | 1208 | 0.29 | 240,000 | <0.1 |
NA, not applicable, ND, not determined.
a
Equilibrium dissociation constants (kd/ka).
b
Dissociation rate constants.
c
Association rate constants.
d
Residence time (RT = 1/(kd)). WT mix: WT protein samples prior and after ATP preincubation were mixed in a 1:1 ratio and immobilized for SPR experiments.