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Updated: Jun 11, 2026
Resin-Assisted Capture Coupled with Isobaric Tandem Mass Tag Labeling for Multiplexed Quantification of Protein Thiol Oxidation
Published on: June 21, 2021
A comprehensive study on molecular features, immunological expressions, and redox-regulatory functions of
H P U M Harishchandra1, M A H Dilshan1, W K M Omeka2
1Department of Marine Life Sciences & Center for Genomic Selection in Korean Aquaculture, Jeju National University, Jeju, 63243, Republic of Korea.
Abstract:
Glutaredoxins, which belong to the thioredoxin superfamily, are heat-stable redox switches crucial for cellular pathways that respond to oxidative stress as oxidoreductases. This study was conducted to characterize glutaredoxin 3 from Scomber japonicus (SjGrx3) using in silico techniques and diverse functional assays. The SjGrx3 domain architecture comprises a thioredoxin domain and two Grx family domains. Pairwise comparison of SjGrx3 with other orthologs revealed the highest identity (92.0%) and similarity (95.4%) with Grx3 from Thunnus maccoyii. SjGrx3 spatial and temporal expression analyses were performed to determine tissue-specific distribution and immunological involvement during pathogenic invasion. The highest SjGrx3 expression was observed in the brain and heart tissues. Furthermore, significant modulation of SjGrx3 expression was observed in blood and gill tissues upon immunostimulation with polyinosinic: polycytidylic acid, lipopolysaccharides, Vibrio harveyi, and Streptococcus iniae. The recombinant SjGrx3 (rSjGrx3) protein exhibited significant oxygen radical absorbance capacity, bis (2-hydroxyethyl) disulfide reduction activity, and dehydroascorbic reductase activity. Subcellular localization assay confirmed the cytoplasmic and nuclear localization of GFP-tagged SjGrx3 protein. Cellular assays further demonstrated that SjGrx3 enhances cell viability and reduces apoptosis under H2O2-induced oxidative stress. In addition, ferrozine assays indicated that SjGrx3 is involved in maintaining cellular iron homeostasis. Overexpression of SjGrx3 also promoted the wound healing effect in an in vitro cell scratch assay. Collectively, these findings suggest that SjGrx3 plays multiple cellular roles in mitigating oxidative damage and contributes to maintain redox homeostasis during stress response and immune regulation in chub mackerel.
