Cloning and functional characterisation of a peroxiredoxin 1 (NKEF A) cDNA from Atlantic salmon (Salmo salar) and its expression in fish infected with Neoparamoeba perurans

Hsi-Mei Loo, Drew Sutton, Kathryn Schuller

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    29 Citations (Scopus)

    Abstract

    Peroxiredoxin 1 (Prx 1), also known as natural killer enhancing factor A (NKEF A), has been implicated in the immune response of both mammals and fish. Amoebic gill disease (AGD), caused by Neoparamoeba perurans, is a significant problem for the Atlantic salmon (Salmo salar L.) aquaculture industry based in Tasmania, Australia. Here we have cloned and functionally characterized a Prx 1 open reading frame (ORF) from Atlantic salmon liver and shown that Prx 1 gene expression was down-regulated in the gills of Atlantic salmon displaying symptoms of AGD. The Prx 1 ORF encoded all of the residues and motifs characteristic of typical 2-Cys Prx proteins from eukaryotes and the recombinant protein expressed in Escherichia coli catalyzed thioredoxin (Trx)-dependent reduction of H 2O 2, cumene hydroperoxide (CuOOH) and t-butyl hydroperoxide (t-bOOH) with K m values of 122, 77 and 91 μM, respectively, confirming that it was a genuine 2-Cys Prx. The recombinant protein also displayed a double displacement reaction mechanism and a catalytic efficiency (k cat/K m) with H 2O 2 of 1.5 × 10 5 M -1 s -1 which was consistent with previous reports for the 2-Cys Prx family of proteins. This is the first time that a Prx 1 protein has been functionally characterized from any fish species and it paves the way for further investigation of this important 2-Cys Prx family member in fish.

    Original languageEnglish
    Pages (from-to)1074-1082
    Number of pages9
    JournalFish and Shellfish Immunology
    Volume32
    Issue number6
    DOIs
    Publication statusPublished - Jun 2012

    Keywords

    • Amoebic gill disease
    • Atlantic salmon
    • Natural killer enhancing factor
    • Peroxiredoxin

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