FIGURE 5.

Interaction of PAI-1 with immobilized VN and effect of AZ3976. A, sensorgram overlay of the association and dissociation in real time after injection of 10 nm PAI-1 with 10 μm AZ3976 (a), incubated for increasing time intervals (arrow from 0 to 215 min) at 20 °C >900 RU immobilized VN, at change to buffer in the presence of 10 μm AZ3976 (b), and at injection of 20 μl of 25 nm tPA in the presence of 10 μm AZ3976 (c), followed by running buffer (d). The sensorgram of buffer only (dashed line) and reference, 10 nm PAI-1 in buffer without preincubation (marked ref) are included, whereas the sensorgrams of PAI-1 incubated in buffer are not shown for clarity. The ΔRU between the channel with immobilized VN, flow channel 2, and the channel without VN, flow channel 1, is shown. B, data are from the experiment shown in A. Initial binding rate to VN, measured after 10 s, as a function of the incubation time for 10 nm PAI-1 in buffer and for 10 nm PAI-1 with AZ3976 are shown. The lines are fitted to the data assuming single exponential decay, k = 0.0007 (buffer) and 0.01 min⁻¹ (AZ3976), respectively.