Recovery of some functional properties of the detergent-extracted cholinergic receptor protein from Torpedo marmorata after reintegration into a membrane environment.
The change of affinity of the acetylcholine receptor for agonists and the influence of local anaesthetics has been studied in detail in receptor-rich membranes. These properties are changed after solubilisation by ionic detergents. A method for reproducibly reintegrating the receptor protein into a lipid environment is described. Reintegration of the receptor results in partial recovery of the binding and fluorescence properties of the membrane-bound receptor protein. In particular, the slow affinity change caused by agonists can be recovered but not the effect of local anaesthetics on this change. The fluorescence response to cholinergic ligands of the reintegrated receptor protein labelled with quinacrine does not appear identical to that found with the native receptor-rich membranes. It is suggested that the failure to recover the sensitivity to local anaesthetics is at the origin of the difficulties to regain functional reconstitution.
D000818:Animals; D002217:Carbachol / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism*; D002793:Cholic Acids / Q000494:pharmacology; D003840:Deoxycholic Acid / Q000494:pharmacology; D003902:Detergents / Q000494:pharmacology*; D005399:Fishes; D006801:Humans; D010713:Phosphatidylcholines / Q000494:pharmacology; D011318:Prilocaine / Q000494:pharmacology; D011796:Quinacrine; D011950:Receptors, Cholinergic / Q000187:drug effects / Q000378:metabolism*; D014118:Toxins, Biological / Q000494:pharmacology
D002793:Cholic Acids; D003902:Detergents; D010713:Phosphatidylcholines; D011950:Receptors, Cholinergic; D014118:Toxins, Biological; D003840:Deoxycholic Acid; D011318:Prilocaine; D002217:Carbachol; D011796:Quinacrine