Figure 2.

GADD45β_69–86 interacts with recombinant MKK7. Starting at time 0, the indicated concentrations of GADD45β_69–86 were injected for 150 s over SPR sensor surfaces on which 5000 RUs of human recombinant MKK7 had been previously captured by amine coupling. The SPR chip was then washed with buffer alone to monitor ligand dissociation. Fitted sensorgrams show GADD45β_69–86 binding to recombinant MKK7 in RUs. The data were obtained by subtracting the reference channels, containing bovine serum albumin or no polypeptides. The heterogenous ligand equation was the best algorithm to fit the data, suggesting that recombinant MKK7 was immobilized in different orientations on the chip surface. Binding parameters obtained in this manner were as follows: k_on=3.15 × 10² 1/Ms; k_off=1.30 × 10⁻³ 1/s; K_D=3.71 μM