Fig. 1.

Binding of different phosphorylated forms of TH to 14-3-3γ by SPR. 14-3-3γ was immobilized via amine coupling as described under “Experimental Procedures.” TH-pS19 (A) and TH-pS19pS40 (B) were injected at different concentrations (10 nM (black, a), 25 nM (red, b), 50 nM (blue, c), and 100 nM (green, d)), and the association to 14-3-3γ was monitored in terms of response units. C, GST-14-3-3γ was immobilized to a CM5 chip (GE Healthcare) using a GST-immobilization kit (GE Healthcare) according to recommendations by the manufacturer. Sensorgrams were recorded at a flow rate of 30 μl/min, and the figure compares injected (90 μl) nonphosphorylated TH (100 nM, black), TH-pS40 (100 nM, blue), and TH-pS19 (100 nM, red).