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

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• 作者：Grace H. Loo ; Drew L. Sutton ; Kathryn A. Schuller ; ^{kathy.schuller@flinders.edu.au}
• 关键词：Natural killer enhancing factor ; Peroxiredoxin ; Amoebic gill disease ; Atlantic salmon
• 刊名：Fish & Shellfish Immunology
• 出版年：2012
• 期刊代码：115_10504648
• 类别：abs
• 出版时间：June, 2012
• 卷：32
• 期：6
• 页码：1074-1082
• 文件大小：1539 K

摘要

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 H₂O₂, cumene hydroperoxide (CuOOH) and t-butyl hydroperoxide (t-bOOH) with K_m values of 122, 77 and 91聽渭M, 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 H₂O₂ of 1.5聽脳聽10⁵聽M^鈭?聽s^鈭? 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.