title: kappa-Conotoxin PVIIA is a peptide inhibiting the shaker K+ channel.
abstract: kappa-Conotoxin PVIIA (kappa-PVIIA), a 27-amino acid toxin from Conus purpurascens venom that inhibits the Shaker potassium channel, was chemically synthesized in a biologically active form. The disulfide connectivity of the peptide was determined. kappa-Conotoxin PVIIA has the following structure. This is the first Conus peptide known to target K+ channels. [structure: see text] Although the Shaker K+ channel is sensitive to kappa-PVIIA, the rat brain Kv1.1 subtype is resistant. Chimeras between Shaker and the Kv1.1 K+ channels were constructed and expressed in Xenopus oocytes. Only channels containing the putative pore-forming region between the fifth and sixth transmembrane domains of Shaker retained toxin sensitivity, indicating that the toxin target site is in this region of the channel. Evidence is presented that kappa-PVIIA interacts with the external tetraethyl-ammonium binding site on the Shaker channel. Although both kappa-PVIIA and charybdotoxin inhibit the Shaker channel, they must interact differently. The F425G Shaker mutation increases charybdotoxin affinity by 3 orders of magnitude but abolishes kappa-PVIIA sensitivity. The precursor sequence of kappa-PVIIA was deduced from a cDNA clone, revealing a prepropeptide comprising 72 amino acids. The N-terminal region of the kappa-PVIIA prepropeptide exhibits striking homology to the omega-, muO-, and delta-conotoxins. Thus, at least four pharmacologically distinct superfamilies of Conus peptides belong to the same "O" superfamily, with the omega- and kappa-conotoxins forming one branch, and the delta- and muO-conotoxins forming a second major branch.
authors.affiliation: Department of Physiology and Biophysics, Case Western Reserve University, Cleveland, Ohio 44106, USA.
authors.forename: K J
authors.initials: KJ
authors.lastname: Shon
authors.forename: M
authors.initials: M
authors.lastname: Stocker
authors.forename: H
authors.initials: H
authors.lastname: Terlau
authors.forename: W
authors.initials: W
authors.lastname: Stühmer
authors.forename: R
authors.initials: R
authors.lastname: Jacobsen
authors.forename: C
authors.initials: C
authors.lastname: Walker
authors.forename: M
authors.initials: M
authors.lastname: Grilley
authors.forename: M
authors.initials: M
authors.lastname: Watkins
authors.forename: D R
authors.initials: DR
authors.lastname: Hillyard
authors.forename: W R
authors.initials: WR
authors.lastname: Gray
authors.forename: B M
authors.initials: BM
authors.lastname: Olivera
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D001921:Brain / Q000187:drug effects / Q000502:physiology; D018999:Charybdotoxin / Q000494:pharmacology; D020916:Conotoxins; D004220:Disulfides / Q000737:chemistry; D008969:Molecular Sequence Data; D008978:Mollusk Venoms / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D016296:Mutagenesis; D026902:Potassium Channel Blockers; D015221:Potassium Channels / Q000235:genetics / Q000378:metabolism; D011498:Protein Precursors / Q000737:chemistry; D051381:Rats; D017386:Sequence Homology, Amino Acid; D051637:Shaker Superfamily of Potassium Channels; D014982:Xenopus laevis
chemical_list: D020916:Conotoxins; D004220:Disulfides; D008978:Mollusk Venoms; D026902:Potassium Channel Blockers; D015221:Potassium Channels; D011498:Protein Precursors; D051637:Shaker Superfamily of Potassium Channels; C108818:kappa-conotoxin PVIIA; D018999:Charybdotoxin