Exposure of cultured fibroblasts to the peptide PR-11 for the identification of induced proteome alterations and discovery of novel potential ligands.
The PR-11 peptide corresponds to the N-terminal and active region of the endogenously synthesized PR-39 molecule, of porcine origin. It is known to possess various biological effects including antimicrobial properties, angiogenic and anti-inflammatory activities. Apart from its reported activity as a proteasome inhibitor, a more comprehensive understanding of its function, at the molecular level, is still lacking. In this study, we used a label-free shotgun strategy to evaluate the proteomic alterations caused by exposure of cultured fibroblasts to the peptide PR-11. This approach revealed that more than half of the identified molecules were related to signalling, transcription and translation. Proteins directly associated to regulation of angiogenesis and interaction with the hypoxia-inducible factor 1-α (HIF-1α) were significantly altered. In addition, at least three differentially expressed molecules of the NF-κB pathway were detected, suggesting an anti-inflammatory property of PR-11. At last, we demonstrated novel potential ligands of PR-11, through its immobilization for affinity chromatography. Among the eluted molecules, gC1qR, a known complement receptor, appeared markedly enriched. This provided preliminary evidence of a PR-11 ligand possibly involved in the internalization of this peptide. Altogether, our findings contributed to a better understanding of the cellular pathways affected by PR-39 derived molecules.
Programa de Pós-Graduação em Ciências Biológicas, Universidade Federal de Ouro Preto, Ouro Preto, Minas Gerais, Brazil.
Gustavo Silveira
GS
Breguez
Programa de Pós-Graduação em Biotecnologia, Universidade Federal de Ouro Preto, Ouro Preto, Minas Gerais, Brazil.
Leandro Xavier
LX
Neves
Departamento de Farmácia, Universidade Federal de Ouro Preto, Ouro Preto, Minas Gerais, Brazil.
Karina Taciana Santos
KTS
Silva
Programa de Pós-Graduação em Ciências Biológicas, Universidade Federal de Ouro Preto, Ouro Preto, Minas Gerais, Brazil.
Lorran Miranda Andrade
LMA
de Freitas
Programa de Pós-Graduação em Ciências Biológicas, Universidade Federal de Ouro Preto, Ouro Preto, Minas Gerais, Brazil.
Gabriela
G
de Oliveira Faria
Departamento de Ciências Biológicas, Núcleo de Pesquisas em Ciências Biológicas, Universidade Federal de Ouro Preto, Ouro Preto, Minas Gerais, Brazil.
Mauro César
MC
Isoldi
Departamento de Ciências Biológicas, Núcleo de Pesquisas em Ciências Biológicas, Universidade Federal de Ouro Preto, Ouro Preto, Minas Gerais, Brazil.
William
W
Castro-Borges
Departamento de Ciências Biológicas, Núcleo de Pesquisas em Ciências Biológicas, Universidade Federal de Ouro Preto, Ouro Preto, Minas Gerais, Brazil. Electronic address: miltonguerra00@gmail.com.
Milton Hércules Guerra
MHG
de Andrade
D000818:Animals; D023181:Antimicrobial Cationic Peptides / Q000378:metabolism / Q000494:pharmacology*; D002352:Carrier Proteins / Q000378:metabolism; D002478:Cells, Cultured; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000378:metabolism; D055767:Immobilized Proteins / Q000378:metabolism / Q000494:pharmacology; D008024:Ligands; D013058:Mass Spectrometry; D024101:Mitochondrial Proteins / Q000378:metabolism; D016328:NF-kappa B / Q000378:metabolism; D010446:Peptide Fragments / Q000378:metabolism / Q000494:pharmacology; D061988:Proteasome Inhibitors / Q000378:metabolism / Q000494:pharmacology; D020543:Proteome / Q000187:drug effects / Q000378:metabolism; D040901:Proteomics; D051381:Rats; D017208:Rats, Wistar; D013552:Swine
D023181:Antimicrobial Cationic Peptides; C490195:C1qbp protein, rat; D002352:Carrier Proteins; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D055767:Immobilized Proteins; D008024:Ligands; D024101:Mitochondrial Proteins; D016328:NF-kappa B; C501730:PR 11 proline-arginine-rich peptide; D010446:Peptide Fragments; D061988:Proteasome Inhibitors; D020543:Proteome; C072212:PR 39
Label-free shotgun; PR-11; PR-11 binding proteins; PR-39; Proline rich-peptides