title: Differential targeting of nicotinic acetylcholine receptors by novel alphaA-conotoxins.
abstract: We describe the isolation and characterization of two peptide toxins from Conus ermineus venom targeted to nicotinic acetylcholine receptors (nAChRs). The peptide structures have been confirmed by mass spectrometry and chemical synthesis. In contrast to the 12-18 residue, 4 Cys-containing alpha-conotoxins, the new toxins have 30 residues and 6 Cys residues. The toxins, named alphaA-conotoxins EIVA and EIVB, block both Torpedo and mouse alpha1-containing muscle subtype nAChRs expressed in Xenopus oocytes at low nanomolar concentrations. In contrast to alpha-bungarotoxin, alphaA-EIVA is inactive at alpha7-containing nAChRs even at micromolar concentrations. In this regard, alphaA-EIVA is similar to the previously described alpha-conotoxins (e.g. alpha-MI and alpha-GI) which also selectively target alpha1- versus alpha7-containing nAChRs. However, alpha-MI and alpha-GI discriminate between the alpha/delta versus alpha/gamma subunit interfaces of the mouse muscle nAChR with 10,000-fold selectivity. In contrast, alphaA-conotoxin EIVA blocks both the alpha/gamma site and alpha/delta site with equally high affinity but with distinct kinetics. The alphaA-conotoxins thus represent novel probes for the alpha/gamma as well as the alpha/delta binding sites of the nAChR.
authors.affiliation: Department of Biology, University of Utah, Salt Lake City, Utah 84112, USA.
authors.forename: R
authors.initials: R
authors.lastname: Jacobsen
authors.forename: D
authors.initials: D
authors.lastname: Yoshikami
authors.forename: M
authors.initials: M
authors.lastname: Ellison
authors.forename: J
authors.initials: J
authors.lastname: Martinez
authors.forename: W R
authors.initials: WR
authors.lastname: Gray
authors.forename: G E
authors.initials: GE
authors.lastname: Cartier
authors.forename: K J
authors.initials: KJ
authors.lastname: Shon
authors.forename: D R
authors.initials: DR
authors.lastname: Groebe
authors.forename: S N
authors.initials: SN
authors.lastname: Abramson
authors.forename: B M
authors.initials: BM
authors.lastname: Olivera
authors.forename: J M
authors.initials: JM
authors.lastname: McIntosh
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D020916:Conotoxins; D006054:Goldfish; D007700:Kinetics; D051379:Mice; D008969:Molecular Sequence Data; D008978:Mollusk Venoms / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology*; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000302:isolation & purification / Q000494:pharmacology; D011978:Receptors, Nicotinic / Q000187:drug effects* / Q000378:metabolism; D014981:Xenopus
chemical_list: D020916:Conotoxins; D008978:Mollusk Venoms; D010446:Peptide Fragments; D011978:Receptors, Nicotinic; C108141:conotoxin EIVA; C108142:conotoxin EIVB