Biacore analysis with stabilized G-protein-coupled receptors

Using stabilized forms of β1 adrenergic and A2A adenosine G-protein-coupled receptors, we applied Biacore to monitor receptor activity and characterize binding constants of small-molecule antagonists spanning >20,000 fold in affinity. We also illustrate an improved method for tethering His-tagged receptors on NTA chips to yield stable, high-capacity, high-activity surfaces, as well as a novel approach to regenerate receptor-binding sites. Based on our success with this approach, we expect that the combination of stabilized receptors with biosensor technology will become a common method for characterizing members of this receptor family.