Functional characterization of peroxiredoxin 5 from yellowtail clownfish (Amphiprion clarkii): Immunological expression assessment, antioxidant activities, heavy metal detoxification, and nitrosative stress mitigation

IF 2.7 3区 农林科学 Q1 FISHERIES Developmental and comparative immunology Pub Date : 2024-11-12 DOI:10.1016/j.dci.2024.105289
D.C.G. Rodrigo , H.M.V. Udayantha , D.S. Liyanage , W.K.M. Omeka , Y.K. Kodagoda , H.A.C.R. Hanchapola , M.A.H. Dilshan , G.A.N.P. Ganepola , W.A.D.L.R. Warnakula , Gaeun Kim , Jeongeun Kim , Jihun Lee , Qiang Wan , Jehee Lee
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Abstract

Peroxiredoxin 5 (Prdx5) is the last recognized member of Prdx family. It is a unique, atypical, 2-Cys antioxidant enzyme, protecting cells from death caused by reactive oxygen species (ROS). In this study, the Prdx5 ortholog of Amphiprion clarkii (AcPrdx5) was identified and characterized to explore its specific structural features and functional properties. The open reading frame of AcPrdx5 is 573 bp long and encodes 190 amino acids containing a mitochondrial targeting sequence, thioredoxin domain, and two conserved cysteine residues responsible for antioxidant function. The predicted molecular weight and theoretical isoelectric point of AcPrdx5 are 20.3 kDa and 9.01, respectively. AcPrdx5 sequences were found to be highly conserved across the other orthologs from various organisms and it distinctively clustered within the fish Prdx5 subclade of the phylogenetic tree. The expression of AcPrdx5 was ubiquitously detected among twelve tested tissues, with the highest level in the brain. Furthermore, the mRNA levels of AcPrdx5 in the blood and head-kidney tissues were significantly (p < 0.05) upregulated following polyinosinic-polycytidylic acid (Poly I:C), lipopolysaccharide (LPS), and Vibrio harveyi immune challenge. A concentration-dependent antioxidant potential of recombinant AcPrdx5 was observed in insulin disulfide bond reduction, heavy metal detoxification, free radical and hydrogen peroxide (H2O2) scavenging assays. Additionally, AcPrdx5 overexpression in fathead minnow (FHM) cells upregulated the antioxidant-associated gene (Rrm1, MAPK, SOD2, and PRDX1) expression after H2O2 treatment, and promoted cell viability upon arsenic (As) exposure. In macrophages, AcPrdx5 overexpression effectively suppressed substantial nitric oxide production under lipopolysaccharide treatment. Collectively, our results suggest that AcPrdx5 may play roles in both antioxidant defense system and innate immune response against pathogenic invasions in A. clarkii.
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黄尾小丑鱼(Amphiprion clarkii)过氧化还原酶 5 的功能特征:免疫学表达评估、抗氧化活性、重金属解毒和亚硝酸应激缓解。
过氧化还原酶 5(Prdx5)是 Prdx 家族中最后一个被确认的成员。它是一种独特的非典型 2-Cys 抗氧化酶,能保护细胞免受活性氧(ROS)导致的死亡。本研究鉴定并表征了克氏蛙的 Prdx5 同源物(AcPrdx5),以探索其特定的结构特征和功能特性。AcPrdx5 的开放阅读框长 573 bp,编码 190 个氨基酸,包含线粒体靶向序列、硫氧还蛋白结构域和两个负责抗氧化功能的保守半胱氨酸残基。AcPrdx5 的预测分子量和理论等电点分别为 20.3 kDa 和 9.01。研究发现,AcPrdx5的序列在来自不同生物的其他直向同源物中高度保守,并且在系统发生树的鱼类Prdx5亚支系中明显聚类。AcPrdx5在12个被测组织中均有表达,其中大脑中的表达水平最高。此外,在多聚肌苷酸(Poly I:C)、脂多糖(LPS)和哈维弧菌免疫挑战后,血液和头肾组织中 AcPrdx5 的 mRNA 水平显著上调(p < 0.05)。在胰岛素二硫键还原、重金属解毒、自由基和过氧化氢(H2O2)清除试验中,观察到重组 AcPrdx5 具有浓度依赖性抗氧化潜力。此外,在黑头鲦鱼(FHM)细胞中过表达 AcPrdx5 能在 H2O2 处理后上调抗氧化相关基因(Rrm1、MAPK、SOD2 和 PRDX1)的表达,并在砷(As)暴露后提高细胞活力。在巨噬细胞中,过表达 AcPrdx5 能有效抑制脂多糖处理下一氧化氮的大量产生。总之,我们的研究结果表明,AcPrdx5可能在克氏原螯虾的抗氧化防御系统和抵御病原体入侵的先天免疫反应中发挥作用。
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来源期刊
CiteScore
6.20
自引率
6.90%
发文量
206
审稿时长
49 days
期刊介绍: Developmental and Comparative Immunology (DCI) is an international journal that publishes articles describing original research in all areas of immunology, including comparative aspects of immunity and the evolution and development of the immune system. Manuscripts describing studies of immune systems in both vertebrates and invertebrates are welcome. All levels of immunological investigations are appropriate: organismal, cellular, biochemical and molecular genetics, extending to such fields as aging of the immune system, interaction between the immune and neuroendocrine system and intestinal immunity.
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