title: Divalent cations modulate membrane binding and pore formation of a potent antibiotic peptide analog of alamethicin.
abstract: The Ca(2+) modulation of pore formation (and disaggregation) kinetics of a synthetic analog of alamethicin F50/5 ([l-Glu(OMe)(7,18,19)]), a potent antibiotic peptide, was investigated in situ and in vitro. The in situ experiments consisted in whole-cell recording from isolated retinal rod outer segments (OS), because once blocking the only OS endogenous conductance with saturating light, the current flows entirely through the (exogenous) channels formed by the peptide. The kinetics of current change induced by peptide application and removal (in ∼50ms) on the OS extracellular side was measured in the presence of divalent cations at different concentrations. The in vitro experiments consisted on the divalent cations modulation of [l-Glu(OMe)(7,18,19)] binding to a mimetic OS membrane immobilized on a sensor chip surface, employing surface plasmon resonance spectroscopy (SPR). The presence of even low mM Ca(2+) or Mg(2+) sufficed to increase the [l-Glu(OMe)(7,18,19)] apparent affinity for the mimetic OS membrane up to ∼4-fold, which accelerated the activation of the peptide-induced current in OS by ∼10-fold with respect to low Ca(2+). In situ and in vitro experiments indicate that high concentrations of divalent cations increased also membrane rigidity, contrasting their effect on increasing the pore formation rate.
authors.affiliation: Institute for Maternal and Child Health - IRCCS "Burlo Garofolo" - Trieste, Italy.
authors.forename: Marco
authors.initials: M
authors.lastname: Aquila
authors.forename: Mascia
authors.initials: M
authors.lastname: Benedusi
authors.forename: Karl-Wilhelm
authors.initials: KW
authors.lastname: Koch
authors.forename: Daniele
authors.initials: D
authors.lastname: Dell'Orco
authors.forename: Giorgio
authors.initials: G
authors.lastname: Rispoli
mesh_terms: D000408:Alamethicin / Q000031:analogs & derivatives*; D000818:Animals; D000900:Anti-Bacterial Agents / Q000737:chemistry / Q000378:metabolism*; D001665:Binding Sites / Q000187:drug effects; D002118:Calcium / Q000737:chemistry / Q000494:pharmacology*; D002413:Cations, Divalent / Q000378:metabolism / Q000494:pharmacology*; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism*; D007700:Kinetics; D008274:Magnesium / Q000737:chemistry / Q000494:pharmacology; D009842:Oligopeptides / Q000737:chemistry / Q000378:metabolism*; D011898:Ranidae; D012374:Rod Cell Outer Segment / Q000737:chemistry* / Q000378:metabolism
chemical_list: D000900:Anti-Bacterial Agents; D002413:Cations, Divalent; D009842:Oligopeptides; D000408:Alamethicin; D008274:Magnesium; D002118:Calcium