Table 3.
Binding kinetics of CD27 wild-type (glycosylated) and mutants with immobilized CD70 at the temperature of 37 °C
| Analyte | n | ka (104 M−1s−1) | kd (10−3 s−1) | KD (nM) | Result |
|---|---|---|---|---|---|
| CD27 (WT) | 3 | 3.31 (0.42) | 4.47 (0.43) | 135 (22) | Homogeneous kinetics |
| CD27-R30A | 3 | 1.34 (0.07) | 4.70 (0.34) | 351 (31) | |
| CD27-S83A | 3 | 3.36 (0.36) | 8.54 (0.55) | 254 (32) | |
| CD27-N95A | 3 | 7.25 (0.17) | 7.26 (0.04) | 100 (2) | |
| CD27-K115A | 3 | 3.45 (0.32) | 3.51 (0.04) | 102 (9) | |
| CD27-S70A | 1 | -- | -- | -- | Altered kinetics compared with CD27-WTa |
| CD27-D74A | 1 | -- | -- | -- | |
| CD27-N88A | 1 | -- | -- | -- | |
| CD27-R113A | 1 | -- | -- | -- | |
| CD27-D114E | 1 | -- | -- | -- | |
| CD27-D114R | 1 | -- | -- | -- | |
| CD27-T118E | 1 | -- | -- | -- | |
| CD27-T118R | 3 | -- | -- | -- | |
| CD27-D121A | 1 | -- | -- | -- | |
| CD27-E82R | 1 | -- | -- | -- | No binding |
| CD27-E82A | 1 | -- | -- | -- |
a
Kinetics and affinity parameters were not reported due to weak binding (R113A, D114R, T118E, T118R), heterogeneous kinetics (D74A, N88A, D114E, D121A), or slow association (S70A). Triple independent experiments were performed to measure the binding affinities of mutants with homogeneous kinetic profiles. Standard deviations are shown in parenthesis. The number of replicate measurements is listed as n.