A biophysical study of the interaction of the lipopeptide antibiotic iturin A with aqueous phospholipid bilayers.
Iturin A is a lipopeptide extracted from the culture media of Bacillus subtilis which shows a strong antifungal action. The interaction of iturin A with multilamellar vesicles of dimyristoylphosphatidylcholine (DMPC) induced structures which did not sediment during centrifugation. Electron microscopy after negative staining showed that, at 30 mol%, iturin A/DMPC vesicles were visible but smaller than those formed by pure DMPC. Thermograms of DMPC/iturinA obtained after differential scanning calorimetry, at low concentrations of iturin A, were interpreted as indicating the presence of two laterally separated phases, one formed by pure phospholipid and the other by lipopeptide-phospholipid complexes, these two separated phases being already detected even at low concentrations such as 2 mol%. Fluorescence quenching experiments showed that the D-Tyr residue of the lipopeptide was fully accessible to the aqueous medium, indicating that the polar part of iturin A is located outside of the membrane hydrophobic palisade. It was concluded that the membrane barrier properties are likely to be damaged in the area where the lipid complexes are accumulated, due to structural fluctuations, and this may be one of the bases of its biological activity. Iturin-A was also able to greatly destabilize dielaidoylphosphatidylethanolamine (DEPE) membranes in the fluid form, producing a new structure which had a poor correlation in X-ray diffraction, and in 31P NMR spectroscopy gave rise to a spectrum containing a double isotropic signal. Iturin A was shown to induce DEPE to adopt phases other than H(II) inverted hexagonal, underlining that this lipopeptide is capable of modifying the curvature of the membrane, which may also be important in explaining the tendency of iturin A to create small vesicles and which may be another of the bases of its biological activity.
Grau
A
A
Departamento de Bioquímica y Biología Molecular A, Facultad de Veterinaria, Universidad de Murcia, Spain.
Ortiz
A
A
de Godos
A
A
Gómez-Fernández
J C
JC
D000900:Anti-Bacterial Agents / Q000737:chemistry* / Q000378:metabolism; D000935:Antifungal Agents / Q000737:chemistry*; D055592:Biophysical Phenomena; D001703:Biophysics; D002152:Calorimetry, Differential Scanning; D004134:Dimyristoylphosphatidylcholine / Q000737:chemistry; D007700:Kinetics; D008051:Lipid Bilayers / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy; D008854:Microscopy, Electron; D008956:Models, Chemical; D010455:Peptides; D010456:Peptides, Cyclic; D013050:Spectrometry, Fluorescence; D013696:Temperature; D014961:X-Ray Diffraction
D000900:Anti-Bacterial Agents; D000935:Antifungal Agents; D008051:Lipid Bilayers; D010455:Peptides; D010456:Peptides, Cyclic; C013579:iturin A; D004134:Dimyristoylphosphatidylcholine