A single point mutation confers properties of the muscle-type nicotinic acetylcholine receptor to homomeric alpha7 receptors.
Although the muscle-type and homomeric alpha7-type nicotinic acetylcholine receptors (nAChRs) share many structural features and bind alpha-bungarotoxin with high affinity, several important functional and pharmacological properties distinguish these two major nAChR subtypes. We have shown previously that amino acid sequence in the second transmembrane (TM) domain of the beta subunit is critical for pharmacological distinction between muscle type and heteromeric neuronal (e.g., ganglionic) nAChRs. We tested the hypothesis that homologous substitution of amino acid sequence from the muscle beta1 subunit into the alpha7 subunit would confer specific properties of muscle-type receptors to mutant alpha7 nAChRs. In this study, we show that a single amino acid substitution at the alpha7 TM2 6' position makes both biophysical and pharmacological properties of the mutant receptors resemble those of wild-type muscle nAChR. This mutation produces significant changes in acetylcholine potency and response kinetics, eliminating the characteristic fast desensitization of alpha7 and dramatically reducing divalent ion permeability relative to wild-type alpha7. The TM2 T6'F mutation also produces a profound increase in activation by succinylcholine compared with either wild-type alpha7 or neuronal beta-subunit-containing receptors and the loss of potentiation by 5-hydroxyindole. Thus, the alpha7 TM2 T6'F mutant displays several features that are similar to the muscle nAChR, some of which are not typically thought to be regulated by the pore-lining domain of the receptor.
Placzek
Andon N
AN
Department of Pharmacology and Therapeutics, J. Hills Miller-Health Science Center, University of Florida, Gainesville, FL 32610-0267, USA.
Grassi
Francesca
F
Papke
Thaddeus
T
Meyer
Edwin M
EM
Papke
Roger L
RL
D000109:Acetylcholine / Q000378:metabolism*; D000818:Animals; D001464:Barium / Q000378:metabolism; D002463:Cell Membrane Permeability; D004594:Electrophysiology; D007211:Indoles / Q000494:pharmacology; D007700:Kinetics; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D051379:Mice; D009132:Muscles / Q000378:metabolism* / Q000502:physiology; D018722:Nicotinic Agonists / Q000494:pharmacology; D009865:Oocytes / Q000378:metabolism; D017354:Point Mutation; D051381:Rats; D011978:Receptors, Nicotinic / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D013390:Succinylcholine / Q000494:pharmacology; D014982:Xenopus laevis; D064569:alpha7 Nicotinic Acetylcholine Receptor
C579166:Chrna7 protein, mouse; C579165:Chrna7 protein, rat; D007211:Indoles; D008565:Membrane Proteins; D018722:Nicotinic Agonists; D011978:Receptors, Nicotinic; D064569:alpha7 Nicotinic Acetylcholine Receptor; D001464:Barium; C026785:5-hydroxyindole; D013390:Succinylcholine; D000109:Acetylcholine