title: Characterization and Biological Activities of Ocellatin Peptides from the Skin Secretion of the Frog Leptodactylus pustulatus.
abstract: Eight new peptides were isolated from the skin secretion of the frog Leptodactylus pustulatus and their amino acid sequences determined by de novo sequencing and by cDNA cloning. Structural similarities between them and other antimicrobial peptides from the skin secretion of Leptodactylus genus frogs were found. Ocellatins-PT1 to -PT5 (25 amino acid residues) are amidated at the C-terminus, while ocellatins-PT6 to -PT8 (32 amino acid residues) have free carboxylates. Antimicrobial activity, hemolytic tests, and cytotoxicity against a murine fibroblast cell line were investigated. All peptides, except for ocellatin-PT2, have antimicrobial activity against at least one Gram-negative strain. Ocellatin-PT8 inhibited the growth of Escherichia coli, Staphylococcus aureus, Klebsiella pneumoniae, and Salmonella choleraesuis strains with MICs in the 60-240 μM range. No significant effect was observed in human erythrocytes and in a murine fibroblast cell line after exposure to the peptides at MICs. A comparison between sequences obtained by both direct HPLC-MS de novo sequencing and cDNA cloning demonstrates the secretion of mature peptides derived from a pre-pro-peptide structure.
authors.affiliation: †CENPAT-CONICET, Centro Nacional Patagónico, Consejo Nacional de Investigaciones Científicas y Técnicas, Puerto Madryn, Chubut, Argentina.
authors.forename: Mariela Mirta
authors.initials: MM
authors.lastname: Marani
authors.affiliation: ‡Secretaria de Vigilância em Saúde, Ministério da Saúde, SVS/MS, Brasília, DF, Brazil.
authors.forename: Flávio Santos
authors.initials: FS
authors.lastname: Dourado
authors.affiliation: §Núcleo de Pesquisa em Biodiversidade e Biotecnologia, Biotec, Campus Ministro Reis Velloso, Universidade Federal do Piauí, UFPI, Parnaiba, PI, Brazil.
authors.forename: Patrick Veras
authors.initials: PV
authors.lastname: Quelemes
authors.affiliation: §Núcleo de Pesquisa em Biodiversidade e Biotecnologia, Biotec, Campus Ministro Reis Velloso, Universidade Federal do Piauí, UFPI, Parnaiba, PI, Brazil.
authors.forename: Alyne Rodrigues
authors.initials: AR
authors.lastname: de Araujo
authors.affiliation: §Núcleo de Pesquisa em Biodiversidade e Biotecnologia, Biotec, Campus Ministro Reis Velloso, Universidade Federal do Piauí, UFPI, Parnaiba, PI, Brazil.
authors.forename: Márcia Luana Gomes
authors.initials: ML
authors.lastname: Perfeito
authors.affiliation: ⊥Programa de Pós-Graduação em Biologia Molecular, Departamento de Biologia Molecular, Universidade de Brasília, Brasília, DF, Brazil.
authors.forename: Eder Alves
authors.initials: EA
authors.lastname: Barbosa
authors.affiliation: §Núcleo de Pesquisa em Biodiversidade e Biotecnologia, Biotec, Campus Ministro Reis Velloso, Universidade Federal do Piauí, UFPI, Parnaiba, PI, Brazil.
authors.forename: Leiz Maria Costa
authors.initials: LM
authors.lastname: Véras
authors.affiliation: §Núcleo de Pesquisa em Biodiversidade e Biotecnologia, Biotec, Campus Ministro Reis Velloso, Universidade Federal do Piauí, UFPI, Parnaiba, PI, Brazil.|∥REQUIMTE/LAQV, Instituto Superior de Engenharia do Instituto Politécnico do Porto, Porto, Portugal.
authors.forename: Andreia Luísa Rodrigues
authors.initials: AL
authors.lastname: Coelho
authors.affiliation: §Núcleo de Pesquisa em Biodiversidade e Biotecnologia, Biotec, Campus Ministro Reis Velloso, Universidade Federal do Piauí, UFPI, Parnaiba, PI, Brazil.|∇Programa de Pós-Graduação em Biodiversidade e Biotecnologia, BIONORTE, Universidade Federal do Maranhão, UFMA, São Luís, MA, Brazil.
authors.forename: Etielle Barroso
authors.initials: EB
authors.lastname: Andrade
authors.affiliation: ○UCIBIO, REQUIMTE, Departamento de Química e Bioquímica, Faculdade de Ciências, Universidade do Porto, Porto, Portugal.
authors.forename: Peter
authors.initials: P
authors.lastname: Eaton
authors.affiliation: #Department of Genetics and Morphology, Institute of Biological Sciences, Universidade de Brasília, Brasilia, Brazil.
authors.forename: João Paulo Figueiró
authors.initials: JP
authors.lastname: Longo
authors.affiliation: #Department of Genetics and Morphology, Institute of Biological Sciences, Universidade de Brasília, Brasilia, Brazil.
authors.forename: Ricardo Bentes
authors.initials: RB
authors.lastname: Azevedo
authors.affiliation: ∥REQUIMTE/LAQV, Instituto Superior de Engenharia do Instituto Politécnico do Porto, Porto, Portugal.
authors.forename: Cristina
authors.initials: C
authors.lastname: Delerue-Matos
authors.affiliation: §Núcleo de Pesquisa em Biodiversidade e Biotecnologia, Biotec, Campus Ministro Reis Velloso, Universidade Federal do Piauí, UFPI, Parnaiba, PI, Brazil.
authors.forename: José Roberto S A
authors.initials: JR
authors.lastname: Leite
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D023181:Antimicrobial Cationic Peptides / Q000097:blood / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology*; D004912:Erythrocytes / Q000187:drug effects; D004926:Escherichia coli / Q000187:drug effects; D005347:Fibroblasts / Q000187:drug effects; D006801:Humans; D007711:Klebsiella pneumoniae / Q000187:drug effects; D051379:Mice; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D015394:Molecular Structure; D041681:NIH 3T3 Cells; D011898:Ranidae / Q000235:genetics / Q000378:metabolism*; D012475:Salmonella / Q000187:drug effects; D012867:Skin / Q000378:metabolism*; D013211:Staphylococcus aureus / Q000187:drug effects
chemical_list: D023181:Antimicrobial Cationic Peptides; C500819:ocellatin 1 protein, Leptodactylus ocellatus