Functional properties of the acetylcholine receptor incorporated in model lipid membranes. Differential effects of chain length and head group of phospholipids on receptor affinity states and receptor-mediated ion translocation.
Torpedo acetylcholine receptor was reconstituted into liposomes of pure synthetic lipids in order to study the influence of the lipid environment on affinity state transitions and the ion translocation function of the receptor. A critical concentration of 30 to 40% of cholesteryl hemisuccinate was necessary in liposomes made of cholesteryl hemisuccinate and dimyristoyl phosphatidylcholine to mimic the kinetics of agonist-induced state transitions observed in native membranes. With increasing chain length of the saturated lecithins, a marked increase in carbamylcholine dissociation constants was observed. Substitution by other dimyristoyl phospholipids for dimyristoyl phosphatidylcholine had the same, though quantitatively less pronounced effects. Introduction of unsaturation in the acyl chains reverted the effect of increasing chain length. Unsaturated phosphatidylethanolamines in combination with 28-35 mol% of cholesteryl hemisuccinate was the best lipid mixture for reconstitution of the receptor-gating function. When phosphatidylethanolamine was replaced totally or partially by other phospholipids with the same or different acyl chain composition, a marked decrease of ion transport was apparent, even when similar vesicle size, receptor incorporation, and agonist-induced affinity transitions were obtained. Therefore, the maintenance of the affinity state transitions of the reconstituted receptor is a necessary but not sufficient condition for the manifestation of the ion-gating receptor activity. On the other hand, the more unsaturated the acyl chains of phosphatidylethanolamine are, the higher the response that was observed, suggesting that a critical lipid packing is essential for the ion translocation function of the receptor.
D000818:Animals; D002217:Carbachol / Q000494:pharmacology; D002462:Cell Membrane / Q000378:metabolism; D002788:Cholesterol Esters / Q000494:pharmacology; D004557:Electric Organ / Q000378:metabolism; D007700:Kinetics; D008081:Liposomes; D008854:Microscopy, Electron; D008954:Models, Biological; D010743:Phospholipids / Q000494:pharmacology*; D011950:Receptors, Cholinergic / Q000378:metabolism*; D012964:Sodium / Q000378:metabolism; D013329:Structure-Activity Relationship; D014101:Torpedo
D002788:Cholesterol Esters; D008081:Liposomes; D010743:Phospholipids; D011950:Receptors, Cholinergic; D002217:Carbachol; D012964:Sodium; C013440:cholesteryl succinate