title: Topology of the pore-region of a K+ channel revealed by the NMR-derived structures of scorpion toxins.
abstract: The architecture of the pore-region of a voltage-gated K+ channel, Kv1.3, was probed using four high affinity scorpion toxins as molecular calipers. We established the structural relatedness of these toxins by solving the structures of kaliotoxin and margatoxin and comparing them with the published structure of charybdotoxin; a homology model of noxiustoxin was then developed. Complementary mutagenesis of Kv1.3 and these toxins, combined with electrostatic compliance and thermodynamic mutant cycle analyses, allowed us to identify multiple toxin-channel interactions. Our analyses reveal the existence of a shallow vestibule at the external entrance to the pore. This vestibule is approximately 28-32 A wide at its outer margin, approximately 28-34 A wide at its base, and approximately 4-8 A deep. The pore is 9-14 A wide at its external entrance and tapers to a width of 4-5 A at a depth of approximately 5-7 A from the vestibule. This structural information should directly aid in developing topological models of the pores of related ion channels and facilitate therapeutic drug design.
authors.affiliation: Department of Physiology and Biophysics, University of California, Irvine 92717, USA.
authors.forename: J
authors.initials: J
authors.lastname: Aiyar
authors.forename: J M
authors.initials: JM
authors.lastname: Withka
authors.forename: J P
authors.initials: JP
authors.lastname: Rizzi
authors.forename: D H
authors.initials: DH
authors.lastname: Singleton
authors.forename: G C
authors.initials: GC
authors.lastname: Andrews
authors.forename: W
authors.initials: W
authors.lastname: Lin
authors.forename: J
authors.initials: J
authors.lastname: Boyd
authors.forename: D C
authors.initials: DC
authors.lastname: Hanson
authors.forename: M
authors.initials: M
authors.lastname: Simon
authors.forename: B
authors.initials: B
authors.lastname: Dethlefs
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D018999:Charybdotoxin / Q000737:chemistry; D004553:Electric Conductivity; D004563:Electrochemistry; D015640:Ion Channel Gating; D009682:Magnetic Resonance Spectroscopy; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016296:Mutagenesis; D009498:Neurotoxins / Q000737:chemistry; D015221:Potassium Channels / Q000737:chemistry* / Q000502:physiology; D017434:Protein Structure, Tertiary; D012604:Scorpion Venoms / Q000737:chemistry*; D012996:Solutions; D013816:Thermodynamics
chemical_list: D009498:Neurotoxins; D015221:Potassium Channels; D012604:Scorpion Venoms; D012996:Solutions; C072276:kaliotoxin; D018999:Charybdotoxin; C077910:margatoxin; C052567:noxiustoxin