title: Implication of segment S45 in the permeation pathway of voltage-dependent sodium channels.
abstract: A 34-mer peptide, encompassing the S4 and S45 segments of domain IV of the electric eel voltage-dependent sodium channel, was synthesized in order to test the potential implication of S45 in the gating or permeation pathway. The secondary structure of peptide S4-S45 assessed by circular dichroism was found mainly helical, both in organic solvents and in lipid vesicles, especially negatively-charged ones. The macroscopic conductance properties of neutral and negatively-charged Montal-Mueller planar lipid bilayers doped with S4-S45 were studied and compared with those of S4. With regard to voltage-dependence, the most efficient system was S4-S45 in neutral bilayers. Voltage thresholds for exponential conductance development were found to correlate with the background or "leak" conductance. Assuming that the latter reflects interfacial peptide concentration, the mean apparent number of monomers per conducting aggregate could be estimated to be 3-5. In single-channel experiments, the most probable events had amplitudes of 8 pS and 5 pS in neutral and negatively-charged bilayers respectively. Ionic selectivity under salt gradients conditions, both at macroscopic and single-channel levels, was in favour of sodium ions (PNa/PK = 3). These properties compare favourably to previous reports dealing with peptide modelling transmembrane segments of voltage-dependent ionic channels. Specifically, when compared to S4 alone, the reduced unit conductance and the increased selectivity for sodium support the implication of the S45 region in the inner lining of the open configuration of sodium channels.
authors.affiliation: URA 500 CNRS, Université de Rouen, Mont-Saint-Aignan, France.
authors.forename: M
authors.initials: M
authors.lastname: Brullemans
authors.forename: O
authors.initials: O
authors.lastname: Helluin
authors.forename: J Y
authors.initials: JY
authors.lastname: Dugast
authors.forename: G
authors.initials: G
authors.lastname: Molle
authors.forename: H
authors.initials: H
authors.lastname: Duclohier
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002851:Chromatography, High Pressure Liquid; D002942:Circular Dichroism; D004593:Electrophorus; D004594:Electrophysiology; D066298:In Vitro Techniques; D015640:Ion Channel Gating; D008051:Lipid Bilayers; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000378:metabolism* / Q000502:physiology; D010539:Permeability; D017433:Protein Structure, Secondary; D015222:Sodium Channels / Q000378:metabolism* / Q000502:physiology; D016339:Spectrometry, Mass, Fast Atom Bombardment
chemical_list: D008051:Lipid Bilayers; D010446:Peptide Fragments; D015222:Sodium Channels