Human α 6 β 4 Nicotinic Acetylcholine Receptor: Heterologous Expression and Agonist Behavior Provide Insights into the Immediate Binding Site

• Published in: Molecular pharmacology 2023-06, Vol.103 (6), p.339-347
• Main Authors: Maldifassi, María Constanza, Rego Campello, Hugo, Gallagher, Timothy, Lester, Henry A, Dougherty, Dennis A
• Format: Article
• Language: English
• Subjects: Acetylcholine - metabolism; Acetylcholine - pharmacology; Animals; Binding Sites; Humans; Ligands; Oocytes - metabolism; Protein Domains; Receptors, Nicotinic - genetics; Receptors, Nicotinic - metabolism; Xenopus laevis - metabolism
• ISSN: 0026-895X; 1521-0111
• DOI: 10.1124/molpharm.123.000672

Study of 6 4 nicotinic acetylcholine receptors (nAChRs) as a pharmacological target has recently gained interest because of their involvement in analgesia, control of catecholamine secretion, dopaminergic pathways, and aversive pathways. However, an extensive characterization of the human 6 4 nAChRs has been vitiated by technical difficulties resulting in poor receptor expression. In 2020, Knowland and collaborators identified BARP ( -anchoring and regulatory protein), a previously known voltage-gated calcium channel suppressor, as a novel human 6 4 chaperone. Here, we establish that co-expression of human BARP with human 6 4 in oocytes, resulted in the functional expression of human 6 4 receptors with acetylcholine-elicited currents that allow an in-depth characterization of the receptor using two electrode voltage-clamp electrophysiology together with diverse agonists and receptor mutations. We report: 1) an extended pharmacological characterization of the receptor, and 2) key residues for agonist-activity located in or near the first shell of the binding pocket. SIGNIFICANCE STATEMENT: The human α6β4 nicotinic acetylcholine receptor has attained increased interest because of its involvement in diverse physiological processes and diseases. Although recognized as a pharmacological target, development of specific agonists has been hampered by limited knowledge of its structural characteristics and by challenges in expressing the receptor. By including the chaperone -anchoring and regulatory protein for enhanced expression and employing different ligands, we have studied the pharmacology of α6β4, providing insight into receptor residues and structural requirements for ligands important to consider for agonist-induced activation.