Interaction of the lantibiotic nisin with membranes revealed by fluorescence quenching of an introduced tryptophan.
Nisin is a lantibiotic produced by strains of Lactococcus lactis subsp. lactis. The target for nisin action is the cytoplasmic membrane of gram-positive bacteria. To aid understanding of its mode of action, the interaction of nisin with vesicles of differing phospholipid composition were investigated by fluorescence techniques, using a variant of nisin in which the isoleucine at position 30 was replaced by a tryptophan residue. Activity of the site-directed variant containing tryptophan was established to be similar to that of the wild-type peptide. Fluorescence experiments showed a blue shift of the emission wavelength maximum in the presence of lipid vesicles, indicating that the tryptophan residue enters a more hydrophobic environment. Quenching experiments with aqueous and membrane-restricted quenchers (iodide and spin-labelled lipids, respectively) both confirmed a non-aqueous environment for the Trp30 residue, and implied that the residue resides between 0.36 nm and 0.52 nm from the centre of the membrane, depending on the lipid identity. The results clearly demonstrate that nisin interacts strongly with the hydrophobic phase of lipid vesicles. This interaction is stronger in the presence of negatively charged lipids suggesting their importance in the functional interaction of nisin with membranes.
Martin
I
I
Laboratoire de Chimie-Physique des Macromolécules aux Interfaces CP206/2, Université Libre de Bruxelles, Belgium.
Ruysschaert
J M
JM
Sanders
D
D
Giffard
C J
CJ
D000595:Amino Acid Sequence; D000900:Anti-Bacterial Agents / Q000737:chemistry*; D002462:Cell Membrane / Q000737:chemistry; D008051:Lipid Bilayers / Q000737:chemistry*; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009561:Nisin / Q000737:chemistry* / Q000235:genetics; D010743:Phospholipids / Q000737:chemistry; D013050:Spectrometry, Fluorescence; D013113:Spin Labels; D014364:Tryptophan / Q000235:genetics*
D000900:Anti-Bacterial Agents; D008051:Lipid Bilayers; D010743:Phospholipids; D013113:Spin Labels; D009561:Nisin; D014364:Tryptophan