title: Dermaseptins from Phyllomedusa oreades and Phyllomedusa distincta. Anti-Trypanosoma cruzi activity without cytotoxicity to mammalian cells.
abstract: Amphibian skin secretions are known as a rich source of biologically active molecules, most of which are alkaloids, biogenic amines, and peptides. Dermaseptins are a class of antimicrobial peptides present in tree frogs of the Phyllomedusa genus. They are cationic molecules of 28-34 residues that permeabilize the membrane of Gram-positive and Gram-negative bacteria, yeasts, and filamentous fungi, showing little or no hemolytic activity. This work reports the isolation, molecular mass analysis, primary structure determination, biological activities, and potential therapeutic applications of an antimicrobial peptide found in the skin secretion of Phyllomedusa oreades, which is a newly described amphibian species endemic of the Brazilian savanna. DS 01 is a 29-residue-long peptide with a molecular mass of 2793.39 Da showing antibacterial properties against Gram-positive and Gram-negative bacteria in the range of 3-25 microm. Anti-protozoan activity was investigated using T. cruzi in its trypomatigote and epimastigote forms cultivated in both cell culture and blood media. Within 2 h after incubation with DS 01 at a final concentration of approximately 6 microm, no protozoan cells were detected. Two synthetic dermaseptins, described previously by our group and named dermadistinctins K and L (DD K and DD L), also had their anti-Trypanosoma cruzi activity investigated and demonstrated similar properties. Toxicity of DS 01 to mouse erythrocytes and white blood cells was evaluated by means of atomic force microscopy and flow cytometry. No morphological alterations were observed at a lytic concentration of DS 01, suggesting its therapeutic value especially as an anti-T. cruzi agent to prevent infections during blood transfusion.
authors.affiliation: Laboratório de Espectrometria de Massa, Empresa Brasileira de Pesquisa Agropecuária (EMBRAPA) - Recursos Genéticos e Biotecnologia, Estação Parque Biológico, Final W5, Asa Norte, Brasilia, DF, 70770-900 Brazil.
authors.forename: Guilherme D
authors.initials: GD
authors.lastname: Brand
authors.forename: José Roberto S A
authors.initials: JR
authors.lastname: Leite
authors.forename: Luciano P
authors.initials: LP
authors.lastname: Silva
authors.forename: Sérgio
authors.initials: S
authors.lastname: Albuquerque
authors.forename: Maura V
authors.initials: MV
authors.lastname: Prates
authors.forename: Ricardo B
authors.initials: RB
authors.lastname: Azevedo
authors.forename: Vanessa
authors.initials: V
authors.lastname: Carregaro
authors.forename: João S
authors.initials: JS
authors.lastname: Silva
authors.forename: Vanuza C L
authors.initials: VC
authors.lastname: Sá
authors.forename: Reuber A
authors.initials: RA
authors.lastname: Brandão
authors.forename: Carlos
authors.initials: C
authors.lastname: Bloch
mesh_terms: D000595:Amino Acid Sequence; D029845:Amphibian Proteins; D000818:Animals; D023181:Antimicrobial Cationic Peptides / Q000737:chemistry* / Q000235:genetics* / Q000494:pharmacology*; D002025:Bufonidae / Q000378:metabolism*; D002412:Cations; D002851:Chromatography, High Pressure Liquid; D004305:Dose-Response Relationship, Drug; D004912:Erythrocytes / Q000378:metabolism / Q000648:ultrastructure; D005434:Flow Cytometry; D018625:Microscopy, Atomic Force; D008969:Molecular Sequence Data; D017830:Octoxynol / Q000494:pharmacology; D010455:Peptides / Q000737:chemistry; D017433:Protein Structure, Secondary; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013997:Time Factors; D014344:Trypanocidal Agents / Q000494:pharmacology*; D014349:Trypanosoma cruzi / Q000187:drug effects / Q000378:metabolism*
chemical_list: D029845:Amphibian Proteins; D023181:Antimicrobial Cationic Peptides; D002412:Cations; D010455:Peptides; D014344:Trypanocidal Agents; C070905:dermaseptin; D017830:Octoxynol