title: A single charged surface residue modifies the activity of ikitoxin, a beta-type Na+ channel toxin from Parabuthus transvaalicus.
abstract: We previously purified and characterized a peptide toxin, birtoxin, from the South African scorpion Parabuthus transvaalicus. Birtoxin is a 58-residue, long chain neurotoxin that has a unique three disulfide-bridged structure. Here we report the isolation and characterization of ikitoxin, a peptide toxin with a single residue difference, and a markedly reduced biological activity, from birtoxin. Bioassays on mice showed that high doses of ikitoxin induce unprovoked jumps, whereas birtoxin induces jumps at a 1000-fold lower concentration. Both toxins are active against mice when administered intracerebroventricularly. Mass determination indicated an apparent mass of 6615 Da for ikitoxin vs. 6543 Da for birtoxin. Amino acid sequence determination revealed that the amino-acid sequence of ikitoxin differs from birtoxin by a single residue change from glycine to glutamic acid at position 23, consistent with the apparent mass difference of 72 Da. This single-residue difference renders ikitoxin much less effective in producing the same behavioral effect as low concentrations of birtoxin. Electrophysiological measurements showed that birtoxin and ikitoxin can be classified as beta group toxins for voltage-gated Na+ channels of central neurons. It is our conclusion that the N-terminal loop preceding the alpha-helix in scorpion toxins is one of the determinative domains in the interaction of toxins with the target ion channel.
authors.affiliation: Department of Entomology and Cancer Research Center, Section of Neurobiology, Physiology and Behavior, and Department of Chemistry and Superfund Analytical Laboratory, University of California, Davis, CA, USA.
authors.forename: A Bora
authors.initials: AB
authors.lastname: Inceoglu
authors.forename: Yuki
authors.initials: Y
authors.lastname: Hayashida
authors.forename: Jozsef
authors.initials: J
authors.lastname: Lango
authors.forename: Andrew T
authors.initials: AT
authors.lastname: Ishida
authors.forename: Bruce D
authors.initials: BD
authors.lastname: Hammock
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D004305:Dose-Response Relationship, Drug; D004594:Electrophysiology; D007477:Ions; D013058:Mass Spectrometry; D051379:Mice; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009474:Neurons / Q000187:drug effects; D010455:Peptides / Q000737:chemistry; D017434:Protein Structure, Tertiary; D012604:Scorpion Venoms / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology; D012605:Scorpions / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D015222:Sodium Channels / Q000378:metabolism*; D013997:Time Factors; D014466:Ultraviolet Rays
chemical_list: D007477:Ions; D010455:Peptides; D012604:Scorpion Venoms; D015222:Sodium Channels; C440191:birtoxin; C470095:ikitoxin protein, Parabuthus transvaalicus