title: The influence of allosteric modulators and transmembrane mutations on desensitisation and activation of α7 nicotinic acetylcholine receptors.
abstract: Acetylcholine activates nicotinic acetylcholine receptors (nAChRs) by binding at an extracellular orthosteric site. Previous studies have described several positive allosteric modulators (PAMs) that are selective for homomeric α7 nAChRs. These include type I PAMs, which exert little or no effect on the rate of receptor desensitisation, and type II PAMs, which cause a dramatic loss of agonist-induced desensitisation. Here we report evidence that transmembrane mutations in α7 nAChRs have diverse effects on receptor activation and desensitisation by allosteric ligands. It has been reported previously that the L247T mutation, located toward the middle of the second transmembrane domain (at the 9' position), confers reduced levels of desensitisation. In contrast, the M260L mutation, located higher up in the TM2 domain (at the 22' position), does not show any difference in desensitisation compared to wild-type receptors. We have found that in receptors containing the L247T mutation, both type I PAMs and type II PAMs are converted into non-desensitising agonists. In contrast, in receptors containing the M260L mutation, this effect is seen only with type II PAMs. These findings, indicating that the M260L mutation has a selective effect on type II PAMs, have been confirmed both with previously described PAMs and also with a series of novel α7-selective PAMs. The novel PAMs examined in this study have close chemical similarity but diverse pharmacological properties. For example, they include compounds displaying effects on receptor desensitisation that are typical of classical type I and type II PAMs but, in addition, they include compounds with intermediate properties.
authors.affiliation: Department of Neuroscience, Physiology & Pharmacology, University College London, London, United Kingdom.
authors.forename: Anna
authors.initials: A
authors.lastname: Chatzidaki
authors.affiliation: Department of Chemistry, University College London, London, United Kingdom.
authors.forename: Jarryl M
authors.initials: JM
authors.lastname: D'Oyley
authors.affiliation: Department of Neuroscience, Physiology & Pharmacology, University College London, London, United Kingdom.
authors.forename: JasKiran K
authors.initials: JK
authors.lastname: Gill-Thind
authors.affiliation: Department of Chemistry, University College London, London, United Kingdom.
authors.forename: Tom D
authors.initials: TD
authors.lastname: Sheppard
authors.affiliation: Department of Neuroscience, Physiology & Pharmacology, University College London, London, United Kingdom. Electronic address: n.millar@ucl.ac.uk.
authors.forename: Neil S
authors.initials: NS
authors.lastname: Millar
mesh_terms: D000109:Acetylcholine / Q000494:pharmacology; D000495:Allosteric Site / Q000235:genetics / Q000502:physiology; D000818:Animals; D018678:Cholinergic Agents / Q000737:chemistry / Q000494:pharmacology*; D057809:HEK293 Cells; D006801:Humans; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology; D051379:Mice; D015394:Molecular Structure; D009154:Mutation; D009865:Oocytes; D011869:Radioligand Assay; D051381:Rats; D044406:Receptors, Serotonin, 5-HT3 / Q000378:metabolism; D014982:Xenopus laevis; D064569:alpha7 Nicotinic Acetylcholine Receptor / Q000235:genetics* / Q000378:metabolism*
chemical_list: D018678:Cholinergic Agents; D044406:Receptors, Serotonin, 5-HT3; D064569:alpha7 Nicotinic Acetylcholine Receptor; D000109:Acetylcholine
keywords: Allosteric modulation; Ion channel; Nicotinic acetylcholine receptor; Pharmacology