title: The ion-channel activity of longibrachins LGA I and LGB II: effects of pro-2/Ala and gln-18/Glu substitutions on the alamethicin voltage-gated membrane channels.
abstract: Longibrachins LGA I (Ac Aib Ala Aib Ala Aib(5) Ala Gln Aib Val Aib(10) Gly Leu Aib Pro Val(15) Aib Aib Gln Gln Pheol(20), with Aib: alpha-aminoisobutyric acid, pheol: phenylalaninol) and LGB II are two homologous 20-residue long-sequence peptaibols isolated from the fungus Trichoderma longibrachiatum that differ between them by a Gln-18/Glu substitution. They distinguish from alamethicin by a Pro-2 for Ala replacement, which allowed to examine for the first time with natural Aib-containing analogues, the effect of Pro-2 on the ion-channel properties exhibited by alamethicin. The influence of these structural modifications on the voltage-gated ion-channel forming activity of the peptides in planar lipid bilayers were analysed. The general 'barrel-stave' model of ion-channel activity, already described for alamethicin, was preserved with both longibrachins. The negatively charged LGB II promoted higher oligomerisation levels, which could presumably dilute the repulsive effect of the negative Glu ring near the entrance of the channel and resulted in lower lifetimes of the substates, confirming the strong anchor of the peptide C-terminus at the cis-interface. Reduction of the channel lifetimes was observed for the longibrachins, compared to alamethicin. This argues for a better stabilisation of the channels formed by peptaibols having a proline at position 2, which results in better anchoring of the peptide monomer N-terminus at the trans-bilayer interface. Qualitative assays of the temperature dependence on the neutral longibrachin channel properties demonstrated a high increase of channel lifetimes and a markedly reduced voltage-sensitivity when the temperature was decreased, showing that such conditions may allow to study the channel-forming properties of peptides leading to fast current fluctuations.
authors.affiliation: UMR 6522 CNRS, Université de Rouen (IFRMP 23-GDR 790 CNRS), Boulevard M. de Broglie, 76821, Mont-Saint-Aignan, France.
authors.forename: P
authors.initials: P
authors.lastname: Cosette
authors.forename: S
authors.initials: S
authors.lastname: Rebuffat
authors.forename: B
authors.initials: B
authors.lastname: Bodo
authors.forename: G
authors.initials: G
authors.lastname: Molle
mesh_terms: D000408:Alamethicin / Q000737:chemistry / Q000302:isolation & purification*; D004563:Electrochemistry; D007473:Ion Channels / Q000737:chemistry*; D008051:Lipid Bilayers / Q000737:chemistry; D008969:Molecular Sequence Data; D018408:Patch-Clamp Techniques; D010455:Peptides / Q000737:chemistry / Q000302:isolation & purification; D013696:Temperature; D014242:Trichoderma / Q000737:chemistry*
chemical_list: D007473:Ion Channels; D008051:Lipid Bilayers; D010455:Peptides; C402113:longibrachin LGA I; D000408:Alamethicin