title: Single amino acid substitutions in alpha-conotoxin PnIA shift selectivity for subtypes of the mammalian neuronal nicotinic acetylcholine receptor.
abstract: The alpha-conotoxins, a class of nicotinic acetylcholine receptor (nAChR) antagonists, are emerging as important probes of the role played by different nAChR subtypes in cell function and communication. In this study, the native alpha-conotoxins PnIA and PnIB were found to cause concentration-dependent inhibition of the ACh-induced current in all rat parasympathetic neurons examined, with IC(50) values of 14 and 33 nM, and a maximal reduction in current amplitude of 87% and 71%, respectively. The modified alpha-conotoxin [N11S]PnIA reduced the ACh-induced current with an IC(50) value of 375 nM and a maximally effective concentration caused 91% block. [A10L]PnIA was the most potent inhibitor, reducing the ACh-induced current in approximately 80% of neurons, with an IC(50) value of 1.4 nM and 46% maximal block of the total current. The residual current was not inhibited further by alpha-bungarotoxin, but was further reduced by the alpha-conotoxins PnIA or PnIB, and by mecamylamine. (1)H NMR studies indicate that PnIA, PnIB, and the analogues, [A10L]PnIA and [N11S]PnIA, have identical backbone structures. We propose that positions 10 and 11 of PnIA and PnIB influence potency and determine selectivity among alpha7 and other nAChR subtypes, including alpha3beta2 and alpha3beta4. Four distinct components of the nicotinic ACh-induced current in mammalian parasympathetic neurons have been dissected with these conopeptides.
authors.affiliation: Department of Physiology and Pharmacology, University of Queensland, Brisbane, Queensland 4072, Australia.
authors.forename: R C
authors.initials: RC
authors.lastname: Hogg
authors.forename: L P
authors.initials: LP
authors.lastname: Miranda
authors.forename: D J
authors.initials: DJ
authors.lastname: Craik
authors.forename: R J
authors.initials: RJ
authors.lastname: Lewis
authors.forename: P F
authors.initials: PF
authors.lastname: Alewood
authors.forename: D J
authors.initials: DJ
authors.lastname: Adams
mesh_terms: D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000818:Animals; D001665:Binding Sites / Q000235:genetics; D002121:Calcium Channel Blockers / Q000378:metabolism / Q000494:pharmacology; D002478:Cells, Cultured; D020916:Conotoxins / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D008969:Molecular Sequence Data; D009474:Neurons / Q000378:metabolism*; D018733:Nicotinic Antagonists / Q000378:metabolism* / Q000494:pharmacology; D051381:Rats; D011978:Receptors, Nicotinic / Q000378:metabolism*; D015398:Signal Transduction; D013329:Structure-Activity Relationship
chemical_list: D002121:Calcium Channel Blockers; D020916:Conotoxins; D018733:Nicotinic Antagonists; D011978:Receptors, Nicotinic