title: Structure, antimicrobial activities and mode of interaction with membranes of novel [corrected] phylloseptins from the painted-belly leaf frog, Phyllomedusa sauvagii.
abstract: Transcriptomic and peptidomic analysis of skin secretions from the Painted-belly leaf frog Phyllomedusa sauvagii led to the identification of 5 novel phylloseptins (PLS-S2 to -S6) and also of phylloseptin-1 (PSN-1, here renamed PLS-S1), the only member of this family previously isolated in this frog. Synthesis and characterization of these phylloseptins revealed differences in their antimicrobial activities. PLS-S1, -S2, and -S4 (79-95% amino acid sequence identity; net charge  = +2) were highly potent and cidal against Gram-positive bacteria, including multidrug resistant S. aureus strains, and killed the promastigote stage of Leishmania infantum, L. braziliensis and L. major. By contrast, PLS-S3 (95% amino acid identity with PLS-S2; net charge  = +1) and -S5 (net charge  = +2) were found to be almost inactive against bacteria and protozoa. PLS-S6 was not studied as this peptide was closely related to PLS-S1. Differential scanning calorimetry on anionic and zwitterionic multilamellar vesicles combined with circular dichroism spectroscopy and membrane permeabilization assays on bacterial cells indicated that PLS-S1, -S2, and -S4 are structured in an amphipathic α-helix that disrupts the acyl chain packing of anionic lipid bilayers. As a result, regions of two coexisting phases could be formed, one phase rich in peptide and the other lipid-rich. After reaching a threshold peptide concentration, the disruption of lipid packing within the bilayer may lead to local cracks and disintegration of the microbial membrane. Differences in the net charge, α-helical folding propensity, and/or degree of amphipathicity between PLS-S1, -S2 and -S4, and between PLS-S3 and -S5 appear to be responsible for their marked differences in their antimicrobial activities. In addition to the detailed characterization of novel phylloseptins from P. sauvagii, our study provides additional data on the previously isolated PLS-S1 and on the mechanism of action of phylloseptins.
authors: UPMC Univ Paris 06, ER3 Biogenèse des Signaux Peptidiques (BIOSIPE), Paris, France.
authors: Zahid
authors: Z
authors: Raja
authors: Sonia
authors: S
authors: André
authors: Christophe
authors: C
authors: Piesse
authors: Denis
authors: D
authors: Sereno
authors: Pierre
authors: P
authors: Nicolas
authors: Thierry
authors: T
authors: Foulon
authors: Bruno
authors: B
authors: Oury
authors: Ali
authors: A
authors: Ladram
mesh_terms: D000595:Amino Acid Sequence; D029845:Amphibian Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology*; D000818:Animals; D023181:Antimicrobial Cationic Peptides / Q000737:chemistry / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology*; D001001:Anura / Q000378:metabolism*; D001483:Base Sequence; D002463:Cell Membrane Permeability; D003001:Cloning, Molecular; D004305:Dose-Response Relationship, Drug; D004926:Escherichia coli / Q000187:drug effects; D020128:Inhibitory Concentration 50; D007700:Kinetics; D018314:Leishmania infantum / Q000187:drug effects; D008070:Lipopolysaccharides / Q000737:chemistry / Q000378:metabolism; D008566:Membranes / Q000378:metabolism*; D008826:Microbial Sensitivity Tests; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011485:Protein Binding; D011487:Protein Conformation; D012867:Skin / Q000378:metabolism; D013211:Staphylococcus aureus / Q000187:drug effects
chemical_list: D029845:Amphibian Proteins; D023181:Antimicrobial Cationic Peptides; D008070:Lipopolysaccharides
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