Figure 2. Arrestin binding to WT, 4P, and 2P peptides

Peptides were immobilized on the surface of a CM5 sensor chip using methods described in Methods. Arrestin was injected in buffer over the surface for 240 s to measure association, followed by continuous flow of buffer alone to measure dissociation. A: WT, 4P and Scr peptides were immobilized on the surface of the sensor chip at 400 RU. Arrestin (2 μM) was injected over the surface of the sensor chip. The binding of arrestin to Scr peptide was subtracted from arrestin binding to WT and 4P. B: The peptides (2P, 4P, and Scr) were bound to the sensor chip at 200 RU each. 10 μM arrestin was injected over the surface. The binding of arrestin to Scr peptide was subtracted from arrestin binding to 2P and 4P. C: The binding of arrestin (0-50 μM) to 4P and Scr peptides immobilized on the sensor chip surface at 200 RU. The binding of arrestin to Scr is subtracted from the binding to 4P at each concentration of arrestin. These results are from a minimum of four experiments.