title: Ion channel stabilization of synthetic alamethicin analogs by rings of inter-helix H-bonds.
abstract: Rings of inter-helix H-bonds due to Gln at position 7, a highly conserved residue in all pore-forming peptaibols, have been suggested to play an important role in the stabilization of alamethicin channels. In an attempt to test this hypothesis, experimental studies have been undertaken on four synthetic alamethicin non-Aib analogs (Alm-dUL) in which the Gln at position 7 (Q7) is substituted by Ala, Asn, or Ser (Q7A, Q7N, or Q7S). Voltage-dependent pore formation by these analogs in planar lipid bilayers is compared at the macroscopic and single-channel conductance levels. As anticipated, the Q7A substitution abolished all channel-forming activity. The voltage dependence of macroscopic current-voltage curves was conserved with the Q7N substitution but reduced in the Q7S analog. Normalized single-channel conductance ratios between substates follow the same pattern, with the Q7S analog yielding the highest unit conductances. Channel lifetimes were the most significantly modulated parameter with markedly faster kinetics when Gln or Asn was replaced by Ser. The effect of the Q7S substitution on channel lifetimes may be explained through a reduced stabilization of bundles by inter-helix H-bonds.
authors.affiliation: Institut Fédératif de Recherches Multidisciplinaires sur les Peptides, URA 500 CNRS-Université de Rouen, Mont-Saint-Aignan, France. gerard.molle@univ-rouen.fr
authors.forename: G
authors.initials: G
authors.lastname: Molle
authors.forename: J Y
authors.initials: JY
authors.lastname: Dugast
authors.forename: G
authors.initials: G
authors.lastname: Spach
authors.forename: H
authors.initials: H
authors.lastname: Duclohier
mesh_terms: D000408:Alamethicin / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000737:chemistry; D000595:Amino Acid Sequence; D055592:Biophysical Phenomena; D001703:Biophysics; D004553:Electric Conductivity; D006860:Hydrogen Bonding; D007473:Ion Channels / Q000737:chemistry*; D008051:Lipid Bilayers / Q000737:chemistry; D008564:Membrane Potentials; D008969:Molecular Sequence Data; D017433:Protein Structure, Secondary
chemical_list: D007473:Ion Channels; D008051:Lipid Bilayers; D000408:Alamethicin