The antimicrobial peptide trichogin and its interaction with phospholipid membranes.
The interaction of the antimicrobial peptide trichogin GA IV with phospholipid bilayers has been studied. A series of analogs of trichogin was synthesized in which the nitroxide spin label, 4-amino-4-carboxy-2,2,6,6-tetramethylpiperidino-1-oxyl (TOAC), replaced one of the three alpha-aminoisobutyric acid (Aib) residues in the sequence. These modified peptides were used to assess the location of different residues of the peptide in a phospholipid bilayer composed of egg phosphatidylcholine containing 0.4 mol% of a fluorescently labelled phospholipid. We demonstrate that the substitution of Aib residues with TOAC does not alter the manner in which the peptide affects membrane curvature or induces vesicle leakage. The proximity of the nitroxide group on the peptide to the 4,4-difluoro-4-bora-3a,4a-diaza-S-indacene (BODIPY) fluorophore attached to the phospholipid was estimated from the extent of quenching of the fluorescence. By this criterion it was concluded that the peptide penetrates into the bilayer and that Aib4 is the most deeply inserted of the Aib residues. The results suggest that the helix axis of the peptide is oriented along the plane of the membrane. All of the peptides were shown to raise the bilayer to the hexagonal phase transition temperature of dipalmitoleoylphosphatidylethanolamine, indicating that they promote positive membrane curvature. This is a property observed with peptides that do not penetrate deeply into the bilayer or are oriented along the bilayer normal. We also demonstrate trichogin-promoted leakage of the aqueous contents of liposomes. These results indicate that the peptides cause bilayer destabilization. The extent of leakage induced by trichogin is very sensitive to the peptide to lipid ratio over a narrow range.
Epand
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RF
Department of Biochemistry, McMaster University Health Sciences Centre, Hamilton, Ontario, Canada. epand@fhs.csu.mcmaster.ca
Epand
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RM
Monaco
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V
Stoia
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Formaggio
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Crisma
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Toniolo
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D000595:Amino Acid Sequence; D000900:Anti-Bacterial Agents / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D002152:Calorimetry, Differential Scanning; D004578:Electron Spin Resonance Spectroscopy; D006020:Glycopeptides; D066298:In Vitro Techniques; D008051:Lipid Bilayers / Q000737:chemistry*; D055666:Lipopeptides; D008081:Liposomes; D008958:Models, Molecular; D010455:Peptides; D010743:Phospholipids / Q000737:chemistry*; D013050:Spectrometry, Fluorescence; D013113:Spin Labels / Q000138:chemical synthesis; D013329:Structure-Activity Relationship; D013816:Thermodynamics
D000900:Anti-Bacterial Agents; D006020:Glycopeptides; D008051:Lipid Bilayers; D055666:Lipopeptides; D008081:Liposomes; D010455:Peptides; D010743:Phospholipids; D013113:Spin Labels; C116419:trichogin GA IV