title: Maurotoxin and the Kv1.1 channel: voltage-dependent binding upon enantiomerization of the scorpion toxin disulfide bridge Cys31-Cys34.
abstract: Maurotoxin (MTX) is a 34-amino acid polypeptide cross-linked by four disulfide bridges that has been isolated from the venom of the scorpion Scorpio maurus palmatus and characterized. Maurotoxin competed with radiolabeled apamin and kaliotoxin for binding to rat brain synaptosomes and blocked K+ currents from Kv1 channel subtypes expressed in Xenopus oocytes. Structural characterization of the synthetic toxin identified half-cystine pairings at Cys3-Cys24, Cys9-Cys29, Cys13-Cys19 and Cys31-Cys34 This disulfide bridge pattern is unique among known scorpion toxins, particularly the existence of a C-terminal '14-membered disulfide ring' (i.e. cyclic domain 31-34), We therefore studied structure-activity relationships by investigating the structure and pharmacological properties of synthetic MTX peptides either modified at the C-terminus ¿i.e. MTX(1-29), [Abu31,34]-MTX and [Cys31,34, Tyr32]D-MTX) or mimicking the cyclic C-terminal domain [i.e. MTX(31-34)]. Unexpectedly, the absence of a disulfide bridge Cys31-Cys34 in [Abu 31,34]-MTX and MTX(1-29) resulted in MTX-unrelated half-cystine pairings of the three remaining disulfide bridges for the two analogs, which is likely to be responsible for their inactivity against Kv1 channel subtypes. Cyclic MTX(31-34) was also biologically inactive. [Cys31,34, Tyr32]D-MTX, which had a 'native', MTX-related, disulfide bridge organization, but a D-residue-induced reorientation of the C-terminal disulfide bridge, was potent at blocking the Kv1.1 channel. This peptide-induced Kv1.1 blockage was voltage-dependent (a property not observed for MTX), maximal in the low depolarization range and associated with on-rate changes in ligand binding. Thus, the cyclic C-terminal domain of MTX seems to be crucial for recognition of Kv1.3, and to a lesser extent, Kv1.2 channels and it may contribute to the stabilization and strength of the interaction between the toxin and the Kv1.1 channel.
authors.affiliation: Laboratoire de Biochimie, CNRS UMR 6560, IFR Jean Roche, Faculté de Médecine Nord, Marseille, France. lecomte.c@jean-roche.univ-mrs.fr
authors.forename: C
authors.initials: C
authors.lastname: Lecomte
authors.forename: R
authors.initials: R
authors.lastname: Ben Khalifa
authors.forename: M F
authors.initials: MF
authors.lastname: Martin-Eauclaire
authors.forename: R
authors.initials: R
authors.lastname: Kharrat
authors.forename: M
authors.initials: M
authors.lastname: El Ayeb
authors.forename: H
authors.initials: H
authors.lastname: Darbon
authors.forename: H
authors.initials: H
authors.lastname: Rochat
authors.forename: M
authors.initials: M
authors.lastname: Crest
authors.forename: J M
authors.initials: JM
authors.lastname: Sabatier
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001030:Apamin / Q000378:metabolism; D001667:Binding, Competitive; D001921:Brain / Q000187:drug effects; D002851:Chromatography, High Pressure Liquid; D004220:Disulfides / Q000737:chemistry*; D004594:Electrophysiology; D051662:Kv1.1 Potassium Channel; D008969:Molecular Sequence Data; D009498:Neurotoxins / Q000737:chemistry* / Q000494:pharmacology; D009865:Oocytes; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000494:pharmacology; D010456:Peptides, Cyclic / Q000138:chemical synthesis / Q000494:pharmacology; D015221:Potassium Channels / Q000378:metabolism*; D024642:Potassium Channels, Voltage-Gated; D051381:Rats; D012604:Scorpion Venoms / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology; D014981:Xenopus
chemical_list: D004220:Disulfides; D009498:Neurotoxins; D010446:Peptide Fragments; D010456:Peptides, Cyclic; D015221:Potassium Channels; D024642:Potassium Channels, Voltage-Gated; D012604:Scorpion Venoms; C072276:kaliotoxin; C104498:maurotoxin; D051662:Kv1.1 Potassium Channel; D001030:Apamin