title: Inhibition of spiralin processing by the lipopeptide antibiotic globomycin.
abstract: The cyclic lipopeptide globomycin, a specific inhibitor of signal-peptidase II (Lsp A), proved toxic for the mollicute Spiroplasma melliferum with a minimal inhibitory concentration (MIC) in the range 6.25-12.5 microM, about one order of magnitude higher (that is, less efficient) than bee-venom mellitin. SDS-PAGE analysis of cell proteins followed by immunolabeling ("Western blotting") and by crossed immunoelectrophoresis demonstrated that the cleavage of the prespiralin leader peptide was prevented by globomycin. Cell fractionation experiments showed that prespiralin was membrane bound and did not accumulate in the cytoplasm or in the culture medium. Furthermore, the use of the potential-sensitive fluorescent dye 3,3'-dipropyl-2,2'-thiadicarbocyanine iodide (diS-C3-[5]) revealed that, in contrast to valinomycin and mellitin, globomycin up to 30 microM has no effect on the electrical transmembrane potential of S. melliferum. This indicates that the toxicity of globomycin for spiroplasma cells is mainly if not exclusively owing to the inhibition of spiralin processing. Added to previously published data, these results suggest that spiralin and probably other lipoproteins of mollicutes are acylated and membrane targeted by a mechanism involving notably the processing of the prelipoprotein precursor by a type II, globomycin-sensitive signal peptidase.
authors.affiliation: Equipe "Membranes et Osmorégulation," CNRS URA No. 256, Université de Rennes 1, Campus de Beaulieu, 35042 Rennes Cedex, France.
authors.forename: L
authors.initials: L
authors.lastname: Béven
authors.forename: M
authors.initials: M
authors.lastname: Le Hénaff
authors.forename: C
authors.initials: C
authors.lastname: Fontenelle
authors.forename: H
authors.initials: H
authors.lastname: Wróblewski
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology* / Q000633:toxicity; D016282:Aspartic Acid Endopeptidases / Q000037:antagonists & inhibitors; D001425:Bacterial Outer Membrane Proteins / Q000378:metabolism*; D001426:Bacterial Proteins / Q000032:analysis; D015153:Blotting, Western; D003593:Cytoplasm / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D008555:Melitten / Q000494:pharmacology / Q000633:toxicity; D008564:Membrane Potentials / Q000502:physiology; D008566:Membranes / Q000737:chemistry; D010455:Peptides; D011480:Protease Inhibitors / Q000494:pharmacology* / Q000633:toxicity; D021382:Protein Sorting Signals / Q000378:metabolism; D013149:Spiroplasma / Q000737:chemistry / Q000187:drug effects* / Q000254:growth & development; D013997:Time Factors; D014634:Valinomycin / Q000378:metabolism
chemical_list: D000900:Anti-Bacterial Agents; D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D010455:Peptides; D011480:Protease Inhibitors; D021382:Protein Sorting Signals; C028588:spiralin; D014634:Valinomycin; D008555:Melitten; C017339:globomycin; D016282:Aspartic Acid Endopeptidases; C095652:signal peptidase II