title: NMR analysis of interaction of LqhalphaIT scorpion toxin with a peptide corresponding to the D4/S3-S4 loop of insect para voltage-gated sodium channel.
abstract: Voltage-gated sodium channels (Navs) are large transmembrane proteins that initiate action potential in electrically excitable cells. This central role in the nervous system has made them a primary target for a large number of neurotoxins. Scorpion alpha-neurotoxins bind to Navs with high affinity and slow their inactivation, causing a prolonged action potential. Despite the similarity in their mode of action and three-dimensional structure, alpha-toxins exhibit great variations in selectivity toward insect and mammalian Navs, suggesting differences in the binding surfaces of the toxins and the channels. The scorpion alpha-toxin binding site, termed neurotoxin receptor site 3, has been shown to involve the extracellular S3-S4 loop in domain 4 of the alpha-subunit of voltage-gated sodium channels (D4/S3-S4). In this study, the binding site for peptides corresponding to the D4/S3-S4 loop of the para insect Nav was mapped on the highly insecticidal alpha-neurotoxin, LqhalphaIT, from the scorpion Leiurus quinquestriatus hebraeus, by following changes in the toxin amide 1H and 15N chemical shifts upon binding. This analysis suggests that the five-residue turn (residues LqK8-LqC12) of LqhalphaIT and those residues in its vicinity interact with the D4/S3-S4 loop of Nav. Residues LqR18, LqW38, and LqA39 could also form a patch contributing to the interaction with D4/S3-S4. Moreover, a new bioactive residue, LqV13, was identified as being important for Nav binding and specifically for the interaction with the D4/S3-S4 loop. The contribution of LqV13 to NaV binding was further verified by mutagenesis. Future studies involving other extracellular regions of Navs are required for further characterization of the structure of the LqhalphaIT-Navs binding site.
authors.affiliation: Department of Structural Biology, Weizmann Institute of Science, Rehovot 76100, Israel.
authors.forename: Einat
authors.initials: E
authors.lastname: Schnur
authors.forename: Michael
authors.initials: M
authors.lastname: Turkov
authors.forename: Roy
authors.initials: R
authors.lastname: Kahn
authors.forename: Dalia
authors.initials: D
authors.lastname: Gordon
authors.forename: Michael
authors.initials: M
authors.lastname: Gurevitz
authors.forename: Jacob
authors.initials: J
authors.lastname: Anglister
mesh_terms: D019943:Amino Acid Substitution / Q000502:physiology; D000818:Animals; D001665:Binding Sites; D002772:Cholera Toxin / Q000737:chemistry / Q000378:metabolism; D004175:Diptera; D029721:Drosophila Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D004331:Drosophila melanogaster; D004594:Electrophysiology; D006863:Hydrogen-Ion Concentration; D019476:Insect Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D007814:Larva / Q000187:drug effects; D008958:Models, Molecular; D019906:Nuclear Magnetic Resonance, Biomolecular; D009865:Oocytes / Q000187:drug effects / Q000502:physiology; D010455:Peptides / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding / Q000502:physiology; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D012604:Scorpion Venoms / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D012605:Scorpions; D015222:Sodium Channels / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D013696:Temperature; D014982:Xenopus laevis
chemical_list: D029721:Drosophila Proteins; D019476:Insect Proteins; C065190:LqhalphaIT toxin; D010455:Peptides; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D012604:Scorpion Venoms; D015222:Sodium Channels; C469950:para protein, Drosophila; D002772:Cholera Toxin