Acetylcholine receptor. Binding properties and ion permeability response after covalent attachment of the local anaesthetic quinacrine.
Membrane vesicles rich in nicotinic acetylcholine receptor prepared from Torpedo californica electric tissue have been irreversibly modified with quinacrine mustard, an alkylating derivative of the local anaesthetic quinacrine. The reaction blocked the ion channel regulated by the acetylcholine receptor. Acetylcholine still bound to the modified membrane vesicles with KD approx. 10(-8). The number of binding sites was reduced by up to 50%. Stopped-flow experiments showed that in contrast to what had been found with the reversibly binding quinacrine no fluorescence changes caused by energy transfer from the irradiated protein to the fluorescent local anaesthetic occurred after addition of agonist. This indicates that the conformational changes associated with the activation of the ion channel are blocked by the covalent reaction with quinacrine mustard. Analysis of the membrane vesicles by SDS-polyacrylamide gel electrophoresis showed that all polypeptide chains assumed to be part of the receptor complex had reacted with the mustard. Even small components, probably lipids, migrating with the dye front, showed fluorescence.
D000818:Animals; D004557:Electric Organ / Q000378:metabolism*; D004735:Energy Transfer; D005399:Fishes; D008566:Membranes / Q000378:metabolism; D010539:Permeability; D011796:Quinacrine / Q000031:analogs & derivatives*; D011797:Quinacrine Mustard / Q000494:pharmacology*; D011950:Receptors, Cholinergic / Q000187:drug effects / Q000378:metabolism*; D012964:Sodium / Q000378:metabolism; D013050:Spectrometry, Fluorescence
D011950:Receptors, Cholinergic; D011797:Quinacrine Mustard; D012964:Sodium; D011796:Quinacrine