Specific interactions of clausin, a new lantibiotic, with lipid precursors of the bacterial cell wall.
We investigated the specificity of interaction of a new type A lantibiotic, clausin, isolated from Bacillus clausii, with lipid intermediates of bacterial envelope biosynthesis pathways. Isothermal calorimetry and steady-state fluorescence anisotropy (with dansylated derivatives) identified peptidoglycan lipids I and II, embedded in dodecylphosphocholine micelles, as potential targets. Complex formation with dissociation constants of approximately 0.3 muM and stoichiometry of approximately 2:1 peptides/lipid intermediate was observed. The interaction is enthalpy-driven. For the first time, to our knowledge, we evidenced the interaction between a lantibiotic and C(55)-PP-GlcNAc, a lipid intermediate in the biosynthesis of other bacterial cell wall polymers, including teichoic acids. The pyrophosphate moiety of these lipid intermediates was crucial for the interaction because a strong binding with undecaprenyl pyrophosphate, accounting for 80% of the free energy of binding, was observed. No binding occurred with the undecaprenyl phosphate derivative. The pentapeptide and the N-acetylated sugar moieties strengthened the interaction, but their contributions were weaker than that of the pyrophosphate group. The lantibiotic decreased the mobility of the pentapeptide. Clausin did not interact with the water-soluble UDP-MurNAc- and pyrophosphoryl-MurNAc-pentapeptides, pointing out the importance of the hydrocarbon chain of the lipid target.
Bouhss
Ahmed
A
CNRS, UMR 8619, Institut de Biochimie et Biophysique Moléculaire et Cellulaire, Orsay, France. ahmed.bouhss@u-psud.fr
Al-Dabbagh
Bayan
B
Vincent
Michel
M
Odaert
Benoit
B
Aumont-Nicaise
Magalie
M
Bressolier
Philippe
P
Desmadril
Michel
M
Mengin-Lecreulx
Dominique
D
Urdaci
Maria C
MC
Gallay
Jacques
J
D001407:Bacillus / Q000302:isolation & purification / Q000378:metabolism; D001419:Bacteria / Q000378:metabolism*; D001430:Bacteriocins / Q000302:isolation & purification / Q000378:metabolism*; D002151:Calorimetry; D002473:Cell Wall / Q000378:metabolism*; D003619:Dansyl Compounds / Q000378:metabolism; D005453:Fluorescence; D005454:Fluorescence Polarization; D007700:Kinetics; D009005:Monosaccharides / Q000378:metabolism; D009038:Motion; D009842:Oligopeptides / Q000378:metabolism; D011106:Polyisoprenyl Phosphates / Q000378:metabolism; D011485:Protein Binding; D012399:Rotation; D013816:Thermodynamics; D013997:Time Factors; D014538:Uridine Diphosphate N-Acetylmuramic Acid / Q000031:analogs & derivatives / Q000378:metabolism
D001430:Bacteriocins; D003619:Dansyl Compounds; D009005:Monosaccharides; D009842:Oligopeptides; D011106:Polyisoprenyl Phosphates; D014538:Uridine Diphosphate N-Acetylmuramic Acid; C545326:clausin peptide, Bacillus clausii; C438940:lipid I; C069249:muramyl-NAc-(pentapeptide)pyrophosphoryl-undecaprenol; C002225:UDP-N-acetylmuramic acid pentapeptide; C088139:undecaprenyl pyrophosphate