Figure 2

Interaction of R-subunit isoforms with immobilised cAMP analogs investigated by SPR. Binding of all four R-subunit isoforms (100 nM) to immobilised 8-AEA-cAMP (A), 8-AHA-cAMP (B), Sp-8-AEA-cAMPS (C) and Sp-8-AHA-cAMPS (D) Sp-2-AHA-cAMPS (E) using a flow rate of 10 μL/min in buffer A containing 0.005% P20. Association and dissociation of hRIα (blue), hRIβ (red), hRIIα (green) and rRIIβ (black) were monitored for 5 and 10 min, respectively. The dissociation was monitored in the presence of 3 mM cGMP. (F) hRIα binding to five different cAMP analogs. 100 nM hRIα was injected to immobilised 8-AHA-cAMP (red), 8-AEA-cAMP (orange), Sp-2-AHA-cAMPS (green), Sp-8-AEA-cAMPS (blue) and Sp-8-AHA-cAMPS (black). Dissociation was initiated either by buffer A containing 0.005% P20 (dotted lines) or by 3 mM cGMP in the same buffer (solid lines). Experimental setup was performed as described above.