TABLE 3.
SPR results for binding of chemokines to mutant EVM1
| EMV1 variant | Binding parameters for hCCL3a
|
Affinity for hCCL2 (KDb [nM]) | ||||
|---|---|---|---|---|---|---|
| ka (106M−1s−1) | kd (10−4s−1) | KD (pM) | KD (mut)/KD (wt) | t1/2 (min) | ||
| WT | 12 ± 4 | 3.4 ± 0.6 | 29 ± 11 | 34 | 44 ± 2 | |
| E17R | 15 ± 3 | 4 ± 1 | 26 ± 12 | 0.9 | 29 | NDc |
| E60R | 7 ± 1 | 4 ± 1 | 57 ± 18 | 2 | 29 | ND |
| Y69R | 9 ± 1 | 140 ± 20 | 1,500 ± 300 | 52 | 0.8 | >1,000 |
| T84R | 9 ± 1 | 4.2 ± 0.4 | 49 ± 9 | 1.7 | 28 | ND |
| L125R | 22 ± 8 | 4 ± 1 | 18 ± 8 | 0.6 | 29 | ND |
| S134R | 6 ± 1 | 7.0 ± 0.8 | 130 ± 30 | 4 | 17 | 500 ± 30 |
| N136W | 5 ± 1 | 7 ± 1 | 140 ± 40 | 5 | 17 | 240 ± 20 |
| K138Y | 5 ± 1 | 1.3 ± 0.4 | 25 ± 8 | 0.9 | 89 | ND |
| S171W | 18 ± 3 | 130 ± 20 | 720 ± 160 | 24 | 0.9 | >1,000 |
| S171Y | 17 ± 2 | 390 ± 50 | 2,300 ± 400 | 79 | 0.3 | >1,000 |
| I173Y | 12 ± 2 | 310 ± 20 | 2,600 ± 500 | 90 | 0.4 | >1,000 |
| I173R | 3.5 ± 0.7 | 470 ± 60 | 13,000 ± 3,000 | 448 | 0.3 | >1,000 |
| Δ51-65 | 2.2 ± 0.6 | 230 ± 30 | 10,000 ± 3,000 | 344 | 0.5 | >1,000 |
a
Values for ka, kd, and KD are means ± standard deviations. KD = kd/ka; t1/2 = 0.693/kd.
b
KD from equilibrium binding analysis.
c
ND indicates measurements that were not determined.