Table 2.
Kinetics of ligand binding to LRP-1 measured by SPR.
| Ligand | ka (105) M−1 s−1 | kd (10–3) (s-−1) | KD (10–9) (M) |
|---|---|---|---|
| RAP | 68.8 | 4.19 | 0.60 |
| TIMP-3 | 23.1 | 3.89 | 1.68 |
| N-TIMP-3 | 5.67 | 3.21 | 5.65 |
| TIMP-1 | 3.53 | 10.9 | 30.9 |
| TIMP-2 | 1.04 | 2.82 | 27.1 |
| MMP-1 E/A | 1.36 | 4.68 | 34.6 |
| MMP-1 ΔC | No binding | No binding | No binding |
| MMP-1/TIMP-3 | 4.61 | 2.28 | 4.96 |
| MMP-1 ΔC /TIMP-3 | 3.96 | 2.54 | 6.42 |
| MMP-1/TIMP-1 | 3.90 | 6.40 | 16.5 |
| MMP-1/TIMP-2 | 4.19 | 3.27 | 7.80 |
| MMP-1/N-TIMP-3 | 5.73 | 2.46 | 4.30 |
| MMP-13 E/A | 2.34 | 5.98 | 25.6 |
| MMP-13/TIMP-3 | 8.82 | 2.95 | 3.35 |
| MMP-13/TIMP-1 | 3.26 | 3.45 | 10.6 |
| MMP-13/TIMP-2 | 4.31 | 2.41 | 5.59 |
| MMP-14 | No binding | No binding | No binding |
| MMP-14/TIMP-3 | 7.85 | 2.32 | 2.95 |
| MMP-14/TIMP-1 | No binding | No binding | No binding |
| MMP-14/TIMP-2 | No binding | No binding | No binding |
| ADAMTS5-2 | 29.9 | 6.78 | 2.27 |
| ADAMTS5-4 | 6.55 | 5.47 | 8.35 |
| ADAMTS4-2 | 0.49 | 3.81 | 77.56 |
| ADAMTS5-2/TIMP-3 | 4.57 | 2.06 | 4.5 |
| ADAMTS5-4/TIMP-3 | 6.80 | 2.03 | 3.00 |
| ADAMTS4-2/TIMP-3 | 6.59 | 2.17 | 3.30 |
| ADAM-10 | -No binding | No binding | No binding |
| ADAM-17 | No binding | No binding | No binding |
| ADAM-10/TIMP-3 | 4.46 | 3.86 | 8.67 |
| ADAM-17/TIMP-3 | 5.57 | 3.81 | 6.84 |
LRP-1 was immobilized on a CM5 sensor chip and the binding constant of different analytes were determined. Each determination was performed in triplicate at six concentration for each analyte [0–80 nM except ADAMTS4-2 (0–180 nM), BSA, MMP-1ΔC and MMP-14 ectodomain (0–2 μM)]. Binding constants were calculated using the BIOevaluation software.