1. Exposure of cultured fibroblasts to the peptide PR-11 for the identification of induced proteome alterations and discovery of novel potential ligands.
- Author
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Breguez GS, Neves LX, Silva KTS, de Freitas LMA, de Oliveira Faria G, Isoldi MC, Castro-Borges W, and de Andrade MHG
- Subjects
- Animals, Antimicrobial Cationic Peptides metabolism, Carrier Proteins metabolism, Cells, Cultured, Fibroblasts drug effects, Fibroblasts metabolism, Hypoxia-Inducible Factor 1, alpha Subunit metabolism, Immobilized Proteins metabolism, Immobilized Proteins pharmacology, Ligands, Mass Spectrometry, Mitochondrial Proteins metabolism, NF-kappa B metabolism, Peptide Fragments metabolism, Peptide Fragments pharmacology, Proteasome Inhibitors metabolism, Proteasome Inhibitors pharmacology, Proteome drug effects, Proteome metabolism, Proteomics, Rats, Rats, Wistar, Swine, Antimicrobial Cationic Peptides pharmacology
- Abstract
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., (Copyright © 2016 Elsevier B.V. All rights reserved.)
- Published
- 2016
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