Antimicrobial activity of trout hepcidin

Claudio A. Álvarez, Fanny Guzmán, Constanza Cárdenas, SERGIO HERNAN MARSHALL GONZALEZ, LUIS ALBERTO MERCADO VIANCO

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52 Scopus citations

Abstract

Hepcidin is an antimicrobial peptide and a hormone produced mostly the liver. It is a cysteine-rich peptide with a highly conserved β-sheet structure. Recently, we described the hepcidin expression in liver of rainbow trout and its inducibility by iron overloading and lipopolysaccharide (LPS). Thus, in this work, we focused in analyzing the importance of the peptide conformation associated to its oxidative state in the antimicrobial activity. This peptide showed a α-helix conformation in reduced state and the characteristic β-sheet conformation in the oxidized state. Antimicrobial activity assays showed that the oxidized peptide is more effective than the reduced peptide against Escherichia coli and the important salmon fish pathogen Piscirickettsia salmonis. In addition, confocal analysis of P. salmonis culture exposed to trout hepcidin coupled with rhodamine revealed the intracellular location of this peptide and Sytox permeation assay showed that membrane disruption is not the mechanism of its antimicrobial action. Moreover, a conserved ATCUN motif was detected in the N-terminus of this peptide. This sequence has been described as a small metal-binding site that has been implicated in DNA cleavage. In this work we proved that this peptide is able to induce DNA hydrolysis in the presence of ascorbate and CuCl 2 . When the same experiments were carried out using a variant with truncated N-terminus no DNA hydrolysis was observed. Our results suggest that correct folding of hepcidin is required for its antimicrobial activity and most likely the metal-binding site (ATCUN motif) present in its N-terminus is involved in the oxidative damage to macromolecules.

Original languageEnglish
Pages (from-to)93-101
Number of pages9
JournalFish and Shellfish Immunology
Volume41
Issue number1
DOIs
StatePublished - 30 Apr 2014

Keywords

  • ATCUN motif
  • Innate immunity
  • Oxidized peptide
  • Piscirickettsia salmonis

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