Recovery of Some Functional Properties of the Detergent-Extracted Cholinergic Receptor Protein from Torpedo marmorata after Reintegration into a Membrane Environment

Abstract

The change of affinity of the acetylcholine receptor [from electric organ] for agonists and the influence of local anesthetics was studied in detail in receptor-rich membranes. These properties were changed after solubilization by ionic detergents. A method for reproducibly reintegrating the receptor protein into a lipid environment was described. Reintegration of the receptor resulted in partial recovery of the binding and fluorescence properties of the membrane-bound receptor protein. The slow affinity change caused by agonists could be recovered but not the effect of local anesthetics on this change. The fluorescence response to cholinergic ligands of the reintegrated receptor protein labeled with quinacrine did not appear identical to that found with the native receptor-rich membranes. Apparently the failure to recover the sensitivity to local anesthetics is at the origin of the difficulties to regain functional reconstitution.