PURIFICATION AND RECONSTITUTION OF THE DELTA-OPIOID RECEPTOR

The delta opioid receptor has been purified, in an active form, by succinylmorphine affinity chromatography. The receptor was purified partially from bovine frontal cortex and to apparent homogeneity from neuroblastoma x glioma hybrid NG108-15 cells as observed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by silver staining. Antiserum to the purified bovine receptor inhibited ligand binding to membranes and immunoprecipitated a 58 kDa protein from NG108-15 cells. Reconstitution of the receptor with lipids enhanced binding by 9-fold. The 58 kDa band protein after electroelution and reconstitution with lipids also showed specific binding, indicating that the receptor could be renatured even after SDS-PAGE in an appropriate lipid environment.