Membrane permeabilisation and antimycoplasmic activity of the 18-residue peptaibols, trichorzins PA.
The membrane permeabilisation properties of six linear natural 18-residue peptaibols, termed trichorzins PA, have been assessed on liposomes and on mollicutes (trivial name, mycoplasmas), a class of parasitic bacteria characterized by a small genome, the lack of a cell wall, a minute cell size, and the incorporation in their plasma membrane of exogenously supplied cholesterol. The trichorzins PA used in this study (PA II, PA IV-VI, PA VIII, and PA IX) differ between them by amino acid or amino alcohol substitutions at positions 4, 7, and 18, and form slightly amphipathic alpha-helices. They proved bactericidal for mollicutes belonging to the genera Acholeplasma, Mycoplasma, and Spiroplasma, with minimal inhibitory concentrations (3.12</=MICs</=50 microM) generally 2 to 4 fold higher than those of alamethicin F50, a related 20-residue peptide (1.56</=MICs</=12.5 microM). Spiroplasma cells were apparently not protected by the presence of spiralin on their surface. The activities of the six trichorzins PA were not influenced by their sequence variations and no synergistic effect was observed. Consistent with the marginal effect of cholesterol on the incorporation of the trichorzins PA into liposome bilayers, the antibiotic activity was independent of the amount of cholesterol in the membranes of the different mollicutes. The trichorzins PA and alamethicin inhibited the motility of Spiroplasma melliferum, the helical cells being deformed and split into coccoid forms. Membrane potential measurements in Acholeplasma laidlawii and S. melliferum showed that trichorzin PA V and alamethicin F50 very efficiently depolarized the plasma membrane of mollicutes. This was consistent with fluorescence and 23Na NMR measurements on liposomes that revealed the permeabilisation of the lipid bilayer and the nonselective ionophoric activity of the trichorzins PA. These data suggest that the bactericidal activity exhibited by the trichorzins PA on mollicutes is due to the permeabilisation of the plasma membrane.
Béven
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Groupe Membranes et Osmorégulation, UPRES-A CNRS No. 6026, GDR CNRS No. 790, Université de Rennes 1, Campus de Beaulieu, Bâtiment 14, 35042 Rennes Cedex, France.
Duval
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Rebuffat
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Riddell
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Bodo
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Wróblewski
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D000408:Alamethicin / Q000494:pharmacology; D000900:Anti-Bacterial Agents / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology*; D002455:Cell Division / Q000187:drug effects; D002463:Cell Membrane Permeability / Q000187:drug effects*; D002713:Chlorine / Q000378:metabolism; D036341:Intercellular Signaling Peptides and Proteins; D007476:Ionophores / Q000494:pharmacology; D008051:Lipid Bilayers / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy; D008564:Membrane Potentials / Q000187:drug effects; D008826:Microbial Sensitivity Tests; D054848:Peptaibols; D010455:Peptides; D012964:Sodium / Q000378:metabolism; D013149:Spiroplasma / Q000187:drug effects / Q000254:growth & development; D008973:Tenericutes / Q000187:drug effects*
D000900:Anti-Bacterial Agents; D036341:Intercellular Signaling Peptides and Proteins; D007476:Ionophores; D008051:Lipid Bilayers; D054848:Peptaibols; D010455:Peptides; C113419:trichorzin PA II; C113420:trichorzin PA IV; C113452:trichorzin PA V; C113453:trichorzin PA VI; D000408:Alamethicin; D002713:Chlorine; D012964:Sodium