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

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

doi:10.1016/j.dci.2024.105289

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

Dev Comp Immunol. 2024 Nov 11:105289. doi: 10.1016/j.dci.2024.105289. Online ahead of print.

Authors

Affiliations

¹ Department of Marine Life Sciences & Center for Genomic Selection in Korean Aquaculture, Jeju National University, Jeju 63243, Republic of Korea.
² Department of Marine Life Sciences & Center for Genomic Selection in Korean Aquaculture, Jeju National University, Jeju 63243, Republic of Korea; Marine Life Research Institute, Jeju National University, Jeju 63333, Republic of Korea.
³ Department of Marine Life Sciences & Center for Genomic Selection in Korean Aquaculture, Jeju National University, Jeju 63243, Republic of Korea; Marine Life Research Institute, Jeju National University, Jeju 63333, Republic of Korea. Electronic address: jehee@jejunu.ac.kr.

PMID: 39536807
DOI: 10.1016/j.dci.2024.105289

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.

Keywords: Amphiprion clarkii; Antioxidant; Innate immune response; Oxidative stress; Peroxiredoxin 5.