Nicotinic acetylcholine receptors assembled from the alpha7 and beta3 subunits.
Intracellular recordings were performed in voltage-clamped Xenopus oocytes upon injection with a mixture of cDNAs encoding the beta3 and mutant alpha7 (L247Talpha7) neuronal nicotinic acetylcholine receptor (nAChR) subunits. The expressed receptors maintained sensitivity to methyllycaconitine and to alpha-bungarotoxin but exhibited a functional profile strikingly different from that of the homomeric L247Talpha7 receptor. The heteromeric L247Talpha7beta3 nAChR had a lower apparent affinity and a faster rate of desensitization than L247Talpha7 nAChR, exhibited nonlinearity in the I-V relationship, and was inhibited by 5-hydroxytryptamine, much like wild type alpha7 (WTalpha7) nAChR. Single channel recordings in cell-attached mode revealed unitary events with a slope conductance of 19 picosiemens and a lifetime of 5 ms, both values being much smaller than those of the homomeric receptor channel. Upon injection with a mixture of WTalpha7 and beta3 cDNAs, clear evidence was obtained for the plasma membrane assembly of heteromeric nAChRs, although ACh could not activate these receptors. It is concluded that beta3, long believed to be an orphan subunit, readily co-assembles with other subunits to form heteromeric receptors, some of which may be negative regulators of cholinergic function.
Palma
E
E
Istituto Pasteur-Fondazione Cenci Bolognetti and Dipartimento di Medicina Sperimentale Universita' di Roma "La Sapienza", via delle Messi d' Oro 156, 00158 Roma, Italy.
Maggi
L
L
Barabino
B
B
Eusebi
F
F
Ballivet
M
M
D000157:Aconitine / Q000031:analogs & derivatives / Q000378:metabolism; D000818:Animals; D002038:Bungarotoxins / Q000378:metabolism; D002642:Chick Embryo; D016297:Mutagenesis, Site-Directed; D009865:Oocytes / Q000378:metabolism; D011487:Protein Conformation; D011978:Receptors, Nicotinic / Q000737:chemistry / Q000378:metabolism*; D014981:Xenopus
D002038:Bungarotoxins; D011978:Receptors, Nicotinic; C054634:methyllycaconitine; D000157:Aconitine