title: A Novel Vasoactive Proline-Rich Oligopeptide from the Skin Secretion of the Frog Brachycephalus ephippium.
abstract: Proline-rich oligopeptides (PROs) are a large family which comprises the bradykinin-potentiating peptides (BPPs). They inhibit the activity of the angiotensin I-converting enzyme (ACE) and have a typical pyroglutamyl (Pyr)/proline-rich structure at the N- and C-terminus, respectively. Furthermore, PROs decrease blood pressure in animals. In the present study, the isolation and biological characterization of a novel vasoactive BPP isolated from the skin secretion of the frog Brachycephalus ephippium is described. This new PRO, termed BPP-Brachy, has the primary structure WPPPKVSP and the amidated form termed BPP-BrachyNH2 inhibits efficiently ACE in rat serum. In silico molecular modeling and docking studies suggest that BPP-BrachyNH2 is capable of forming a hydrogen bond network as well as multiple van der Waals interactions with the rat ACE, which blocks the access of the substrate to the C-domain active site. Moreover, in rat thoracic aorta BPP-BrachyNH2 induces potent endothelium-dependent vasodilatation with similar magnitude as captopril. In DAF-FM DA-loaded aortic cross sections examined by confocal microscopy, BPP-BrachyNH2 was found to increase the release of nitric oxide (NO). Moreover, BPP-BrachyNH2 was devoid of toxicity in endothelial and smooth muscle cell cultures. In conclusion, the peptide BPP-BrachyNH2 has a novel sequence being the first BPP isolated from the skin secretion of the Brachycephalidae family. This opens for exploring amphibians as a source of new biomolecules. The BPP-BrachyNH2 is devoid of cytotoxicity and elicits endothelium-dependent vasodilatation mediated by NO. These findings open for the possibility of potential application of these peptides in the treatment of endothelial dysfunction and cardiovascular diseases.
authors.affiliation: Núcleo de Pesquisa em Biodiversidade e Biotecnologia-BIOTEC, Campus Ministro Reis Velloso-CMRV, Universidade Federal do Piauí -UFPI, Parnaíba, PI, Brazil.|Laboratório de Farmacologia Cardiovascular-LFC, Núcleo de Pesquisas em Plantas Medicinais-NPPM, Universidade Federal do Piauí -UFPI, Teresina, PI, Brazil.
authors.forename: Daniel Dias Rufino
authors.initials: DD
authors.lastname: Arcanjo
authors.affiliation: Núcleo de Pesquisa em Biodiversidade e Biotecnologia-BIOTEC, Campus Ministro Reis Velloso-CMRV, Universidade Federal do Piauí -UFPI, Parnaíba, PI, Brazil.
authors.forename: Andreanne Gomes
authors.initials: AG
authors.lastname: Vasconcelos
authors.affiliation: Pulmonary and Cardiovascular Pharmacology, Department of Biomedicine, Aarhus University, Aarhus, Denmark.
authors.forename: Simón Gabriel
authors.initials: SG
authors.lastname: Comerma-Steffensen
authors.affiliation: Núcleo de Pesquisa em Biodiversidade e Biotecnologia-BIOTEC, Campus Ministro Reis Velloso-CMRV, Universidade Federal do Piauí -UFPI, Parnaíba, PI, Brazil.
authors.forename: Joilson Ramos
authors.initials: JR
authors.lastname: Jesus
authors.affiliation: Laboratório de Espectrometria de Massa, EMBRAPA Recursos Genéticos e Biotecnologia, Brasília, DF, Brazil.
authors.forename: Luciano Paulino
authors.initials: LP
authors.lastname: Silva
authors.affiliation: Laboratório de Toxinologia, Instituto de Ciências Biológicas-ICB, Universidade de Brasília-UnB, Brasília, DF, Brazil.
authors.forename: Osmindo Rodrigues
authors.initials: OR
authors.lastname: Pires Júnior
authors.affiliation: Departamento de Bioquímica e Imunologia, Faculdade de Medicina de Ribeirão Preto-FMRP, Universidade de São Paulo-USP, Ribeirão Preto, SP, Brazil.
authors.forename: Claudio Miguel
authors.initials: CM
authors.lastname: Costa-Neto
authors.affiliation: Departamento de Bioquímica e Imunologia, Faculdade de Medicina de Ribeirão Preto-FMRP, Universidade de São Paulo-USP, Ribeirão Preto, SP, Brazil.
authors.forename: Eduardo Brandt
authors.initials: EB
authors.lastname: Oliveira
authors.affiliation: Centro de Análises Proteômicas e Bioquímicas-CAPB, Universidade Católica de Brasília-UCB, Brasília, DF, Brazil.
authors.forename: Ludovico
authors.initials: L
authors.lastname: Migliolo
authors.affiliation: Centro de Análises Proteômicas e Bioquímicas-CAPB, Universidade Católica de Brasília-UCB, Brasília, DF, Brazil.
authors.forename: Octávio Luiz
authors.initials: OL
authors.lastname: Franco
authors.affiliation: Universidade de Ribeirão Preto-UNAERP, Curso de Medicina, Avenida Costábile Romano 2201, Ribeirão Preto, SP, Brazil.
authors.forename: Carolina Baraldi Araújo
authors.initials: CB
authors.lastname: Restini
authors.affiliation: Departamento de Física e Química, Faculdade de Ciências Farmacêuticas de Ribeirão Preto-FCFRP, Universidade de São Paulo-USP, Ribeirão Preto, SP, Brazil.
authors.forename: Michele
authors.initials: M
authors.lastname: Paulo
authors.affiliation: Departamento de Física e Química, Faculdade de Ciências Farmacêuticas de Ribeirão Preto-FCFRP, Universidade de São Paulo-USP, Ribeirão Preto, SP, Brazil.
authors.forename: Lusiane Maria
authors.initials: LM
authors.lastname: Bendhack
authors.affiliation: Laboratório de Espectrometria de Massa, EMBRAPA Recursos Genéticos e Biotecnologia, Brasília, DF, Brazil.
authors.forename: Marcelo Porto
authors.initials: MP
authors.lastname: Bemquerer
authors.affiliation: Laboratório de Farmacologia Cardiovascular-LFC, Núcleo de Pesquisas em Plantas Medicinais-NPPM, Universidade Federal do Piauí -UFPI, Teresina, PI, Brazil.
authors.forename: Aldeidia Pereira
authors.initials: AP
authors.lastname: Oliveira
authors.affiliation: Pulmonary and Cardiovascular Pharmacology, Department of Biomedicine, Aarhus University, Aarhus, Denmark.
authors.forename: Ulf
authors.initials: U
authors.lastname: Simonsen
authors.affiliation: Núcleo de Pesquisa em Biodiversidade e Biotecnologia-BIOTEC, Campus Ministro Reis Velloso-CMRV, Universidade Federal do Piauí -UFPI, Parnaíba, PI, Brazil.
authors.forename: José Roberto de Souza de Almeida
authors.initials: JR
authors.lastname: Leite
mesh_terms: D000595:Amino Acid Sequence; D000806:Angiotensin-Converting Enzyme Inhibitors / Q000138:chemical synthesis / Q000378:metabolism* / Q000494:pharmacology; D000818:Animals; D001001:Anura / Q000378:metabolism*; D001013:Aorta, Thoracic / Q000166:cytology; D020134:Catalytic Domain; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D008297:Male; D009569:Nitric Oxide / Q000378:metabolism; D009842:Oligopeptides / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D007703:Peptidyl-Dipeptidase A / Q000737:chemistry / Q000378:metabolism; D011392:Proline / Q000737:chemistry; D011485:Protein Binding; D017433:Protein Structure, Secondary; D051381:Rats; D017208:Rats, Wistar; D012867:Skin / Q000378:metabolism*
chemical_list: D000806:Angiotensin-Converting Enzyme Inhibitors; D009842:Oligopeptides; D009569:Nitric Oxide; D011392:Proline; D007703:Peptidyl-Dipeptidase A