The Preparation and Study of Detergent-Solubilized Receptors

Summary

This chapter discussed, from a pragmatic standpoint, some available approaches for preparing and assaying detergent-solubilized preparations of membrane-bound receptors. The guidelines for selection of a detergent and assay for detecting ligand-receptor complexes in solution are primarily empirical in nature. Except in extraordinary circumstances, nearly all methods for assaying binding to detergent-solubilized receptors will disturb the equilibrium of *D + R ⇌ *DR.. However, only the most fortunate investigator has the luxury of worrying about whether equilibrium is being perturbed when the binding assay is terminated. Most investigators simply are seeking a sensitive and reliable assay for detergent-solubilized receptor binding that permits an assessment of whether the solubilized ligand-binding site possesses the specificity characteristic of the membrane-bound receptor. If the detergent-solubilized binding site meets the criteria expected for the physiological receptor of interest, then the optimized assay for detergent-solubilized binding will be useful for identification and characterization of the receptor during subsequent purification and reconstitution protocols.