Figure 2.

SPR sensorgrams and the inhibitory activity of the N-TIMP2_{WT, RGD} and N-TIMP2_HD variants. N-TIMP2_{WT, RGD} (A) and N-TIMP2_HD (B) binding to integrin α_vβ₃ is shown. N-TIMP2_{WT, RGD} (C) and N-TIMP2_HD (D) binding to MMP-14_CAT is shown. Dual binding of N-TIMP2_{WT, RGD} (E) and N-TIMP2_HD (F) to integrin α_vβ₃ and MMP-14_CAT is shown. E and F, variants were allowed to flow over integrin α_vβ₃ immobilized on the chip, followed by MMP-14_CAT (blue) or buffer (control 1, red) injection at 300 s. There was an increase in response units (RU) for the variants after MMP-14_CAT injection, which indicates that these variants bind simultaneously to both receptors. In control 2 (green), a buffer was allowed to flow over the integrin α_vβ₃ immobilized chip, followed by MMP-14 injection at 300 s. In control 3 (yellow), a buffer was allowed to flow over the integrin α_vβ₃ immobilized chip throughout the entire experiment. Substrate degradation velocity at different concentrations of N-TIMP2_{WT, RGD} (0–100 nm) (G) and N-TIMP2_HD (0–0.2 nm) (H) were fitted to Morrison's equation (Equation 2) to obtain K_i values.