title: A spectroscopic study of the membrane interaction of the antimicrobial peptide Pleurocidin.
abstract: The cationic amphipathic alpha-helical antibiotic peptide, pleurocidin, from the winter flounder Pleuronectes americanus associates strongly with anionic membranes where it is able to translocate across the membrane, cause dye leakage from vesicles and induce pore like channel conductance. To investigate the mechanism of pleurocidin antibiotic activity in more detail we have applied a variety of spectroscopic techniques to study the interaction of pleurocidin with model membranes. At neutral pH the peptide inserts into membranes containing anionic lipids and, as shown by proton-decoupled 15N solid-state NMR spectroscopy of macroscopically oriented samples, is aligned parallel to the membrane surface. 2H solid-state NMR spectroscopy of chain deuterated phosphatidylethanolamine (PE) and phosphatidylglycerol (PG) lipids in mixed membranes shows that pleurocidin interacts with both the zwitterionic PE and anionic PG but disrupts the lipid acyl chain order of the anionic PG lipids more effectively. At acidic pH the three histidine residues of pleurocidin become protonated and positively charged which does not alter the membrane disrupting effect nor the location of the peptide in the membrane. The results are interpreted in terms of a structural model for pleurocidin inserted into anionic lipid membranes and the implications of our data are discussed in terms of a general mechanism for the antibiotic activity.
authors.affiliation: Faculté de chimie, University Louis Pasteur/CNRS, LC3-UMR7177 Institut le Bel, Strasbourg, France. jmason@chimie.u-strasbg.fr
authors.forename: A James
authors.initials: AJ
authors.lastname: Mason
authors.forename: I N Husnal
authors.initials: IN
authors.lastname: Chotimah
authors.forename: Philippe
authors.initials: P
authors.lastname: Bertani
authors.forename: Burkhard
authors.initials: B
authors.lastname: Bechinger
mesh_terms: D000595:Amino Acid Sequence; D023181:Antimicrobial Cationic Peptides / Q000737:chemistry*; D029941:Fish Proteins / Q000737:chemistry*; D006639:Histidine / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D008055:Lipids / Q000737:chemistry; D009682:Magnetic Resonance Spectroscopy; D008968:Molecular Conformation; D008969:Molecular Sequence Data; D017433:Protein Structure, Secondary; D013050:Spectrometry, Fluorescence; D013053:Spectrophotometry / Q000379:methods*
chemical_list: D023181:Antimicrobial Cationic Peptides; D029941:Fish Proteins; D008055:Lipids; C105851:pleurocidin; D006639:Histidine