title: Role of lipid II and membrane thickness in the mechanism of action of the lantibiotic bovicin HC5.
abstract: Lantibiotics are antimicrobial peptides produced by Gram-positive bacteria, nisin being the most well-known member. Nisin inhibits peptidoglycan synthesis and forms pores at sensitive membranes upon interaction with lipid II, the essential bacterial cell wall precursor. Bovicin HC5, a bacteriocin produced by Streptococcus bovis HC5, has the putative N-terminal lipid II binding motif, and we investigated the mode of action of bovicin HC5 using both living bacteria and model membranes, with special emphasis on the role of lipid II. Bovicin HC5 showed activity against Staphylococcus cohnii and Staphylococcus warneri, but bovicin HC5 hardly interfered with the membrane potential of S. cohnii. In model membranes, bovicin HC5 was not able to cause carboxyfluorescein release or proton influx from DOPC vesicles containing lipid II. Bovicin HC5 blocked lipid II-dependent pore formation activity of nisin, and a high-affinity interaction with lipid II was observed (apparent binding constant [K(a)] = 3.1 × 10(6) M(-1)), with a 1:1 stoichiometry. In DOPC vesicles containing lipid II, bovicin HC5 was able to assemble with lipid II into a prepore-like structure. Furthermore, we observed pore formation activity of bovicin HC5, which was stimulated by the presence of lipid II, in thin membranes. Moreover, bovicin HC5 induced the segregation of lipid II into domains in giant model membrane vesicles. In conclusion, bovicin HC5 has a primary mode of action similar to that of nisin, but some differences regarding the pore-forming capacity were demonstrated.
authors.affiliation: Departamento de Microbiologia, Universidade Federal de Viçosa, Viçosa, Minas Gerais, Brazil
authors.forename: Aline Dias
authors.initials: AD
authors.lastname: Paiva
authors.forename: Eefjan
authors.initials: E
authors.lastname: Breukink
authors.forename: Hilário Cuquetto
authors.initials: HC
authors.lastname: Mantovani
mesh_terms: D000595:Amino Acid Sequence; D001430:Bacteriocins / Q000737:chemistry* / Q000494:pharmacology*; D002462:Cell Membrane / Q000187:drug effects* / Q000378:metabolism; D018613:Microscopy, Confocal; D008969:Molecular Sequence Data; D009561:Nisin / Q000737:chemistry / Q000494:pharmacology; D010713:Phosphatidylcholines / Q000737:chemistry; D013291:Streptococcus / Q000187:drug effects / Q000378:metabolism; D014538:Uridine Diphosphate N-Acetylmuramic Acid / Q000031:analogs & derivatives* / Q000737:chemistry
chemical_list: D001430:Bacteriocins; D010713:Phosphatidylcholines; D014538:Uridine Diphosphate N-Acetylmuramic Acid; C514465:bovicin HC5, Streptococcus bovis HC5; C069249:muramyl-NAc-(pentapeptide)pyrophosphoryl-undecaprenol; D009561:Nisin; C017251:1,2-oleoylphosphatidylcholine