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Mol Pharmacol. 2017 Mar;91(3):250-262. doi: 10.1124/mol.116.107482. Epub 2017 Jan 9.

Differential Contribution of Subunit Interfaces to α9α10 Nicotinic Acetylcholine Receptor Function.

Author information

1
Instituto de Investigaciones en Ingeniería, Genética y Biología Molecular, Dr Héctor N Torres (J.C.B., I.M., M.M. L., M.M., P.V.P., A.B.E.), Instituto de Química Biológica (P.O.C.), and Instituto de Investigaciones Bioquímicas de Bahía Blanca (J.C., C.B), Consejo Nacional de Investigaciones Científicas y Técnicas, Buenos Aires, Argentina; Department of Neuroscience, Physiology and Pharmacology, University College London, United Kingdom (J.K.G.-T., T.C., N.S.M.); Departamento de Química Biológica Facultad de Ciencias Exactas y Naturales (P.O.C.), and Instituto de Farmacología, Facultad de Medicina (P.V.P., A.B.E.), Universidad de Buenos Aires, Buenos Aires, Argentina; and Departamento de Biología, Bioquímica y Farmacia, Universidad Nacional del Sur, Bahía Blanca, Argentina (J.C., C.B).
2
Instituto de Investigaciones en Ingeniería, Genética y Biología Molecular, Dr Héctor N Torres (J.C.B., I.M., M.M. L., M.M., P.V.P., A.B.E.), Instituto de Química Biológica (P.O.C.), and Instituto de Investigaciones Bioquímicas de Bahía Blanca (J.C., C.B), Consejo Nacional de Investigaciones Científicas y Técnicas, Buenos Aires, Argentina; Department of Neuroscience, Physiology and Pharmacology, University College London, United Kingdom (J.K.G.-T., T.C., N.S.M.); Departamento de Química Biológica Facultad de Ciencias Exactas y Naturales (P.O.C.), and Instituto de Farmacología, Facultad de Medicina (P.V.P., A.B.E.), Universidad de Buenos Aires, Buenos Aires, Argentina; and Departamento de Biología, Bioquímica y Farmacia, Universidad Nacional del Sur, Bahía Blanca, Argentina (J.C., C.B). abelgoyhen@gmail.com elgoyhen@dna.uba.ar.

Abstract

Nicotinic acetylcholine receptors can be assembled from either homomeric or heteromeric pentameric subunit combinations. At the interface of the extracellular domains of adjacent subunits lies the acetylcholine binding site, composed of a principal component provided by one subunit and a complementary component of the adjacent subunit. Compared with neuronal nicotinic acetylcholine cholinergic receptors (nAChRs) assembled from α and β subunits, the α9α10 receptor is an atypical member of the family. It is a heteromeric receptor composed only of α subunits. Whereas mammalian α9 subunits can form functional homomeric α9 receptors, α10 subunits do not generate functional channels when expressed heterologously. Hence, it has been proposed that α10 might serve as a structural subunit, much like a β subunit of heteromeric nAChRs, providing only complementary components to the agonist binding site. Here, we have made use of site-directed mutagenesis to examine the contribution of subunit interface domains to α9α10 receptors by a combination of electrophysiological and radioligand binding studies. Characterization of receptors containing Y190T mutations revealed unexpectedly that both α9 and α10 subunits equally contribute to the principal components of the α9α10 nAChR. In addition, we have shown that the introduction of a W55T mutation impairs receptor binding and function in the rat α9 subunit but not in the α10 subunit, indicating that the contribution of α9 and α10 subunits to complementary components of the ligand-binding site is nonequivalent. We conclude that this asymmetry, which is supported by molecular docking studies, results from adaptive amino acid changes acquired only during the evolution of mammalian α10 subunits.

PMID:
28069778
PMCID:
PMC5325082
DOI:
10.1124/mol.116.107482
[Indexed for MEDLINE]
Free PMC Article

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