Department of Marine Life Sciences & Center for Genomic Selection in Korean Aquaculture, Jeju National University, Jeju, 63243, Republic of Korea.
Department of Marine Life Sciences & Center for Genomic Selection in Korean Aquaculture, Jeju National University, Jeju, 63243, Republic of Korea; Marine Science Institute, Jeju, 63333, Republic of Korea.
Fish Shellfish Immunol. 2023 Oct;141:109009. doi: 10.1016/j.fsi.2023.109009. Epub 2023 Aug 19.
Thioredoxin-like protein 1 (TXNL1) is a redox-active protein belonging to the thioredoxin family, which mainly controls the redox status of cells. The TXNL1 gene from Amphiprion clarkii (AcTXNL1) was obtained from a pre-established transcriptome database. The AcTXNL1 is encoded with 289 amino acids and is predominantly localized in the cytoplasm and nucleus. The TXN domain of AcTXNL1 comprises aCGPC motif with redox-reactive thiol (SH-) groups. The spatial distribution pattern of AcTXNL1 mRNA was examined in different tissues, and the muscle was identified as the highest expressed tissue. AcTXNL1 mRNA levels in the blood and gills were significantly increased in response to different immunostimulants. In vitro antioxidant capacity of the recombinant AcTXNL1 protein (rACTXNL1) was evaluated using the ABTS free radical-scavenging activity assay, cupric ion reducing antioxidant capacity assay, turbidimetric disulfide reduction assay, and DNA nicking protection assay. The potent antioxidant activity of rAcTXNL1 exhibited a concentration-dependent manner in all assays. Furthermore, in the cellular environment, overexpression of AcTXNL1 increased cell viability under HO stress and reduced nitric oxide (NO) production induced by lipopolysaccharides (LPS). Collectively, the experimental results revealed that AcTXNL1 is an antioxidant and immunologically important gene in A. clarkii.
硫氧还蛋白样蛋白 1(TXNL1)是一种属于硫氧还蛋白家族的氧化还原活性蛋白,主要控制细胞的氧化还原状态。我们从已建立的转录组数据库中获得了双锯鱼(Amphiprion clarkii)的 TXNL1 基因(AcTXNL1)。AcTXNL1 编码 289 个氨基酸,主要定位于细胞质和细胞核。AcTXNL1 的 TXN 结构域包含具有氧化还原活性硫醇(SH-)基团的 aCGPC 基序。我们检测了 AcTXNL1 mRNA 在不同组织中的空间分布模式,结果发现肌肉是表达量最高的组织。血液和鳃中的 AcTXNL1 mRNA 水平在受到不同免疫刺激剂作用后显著增加。我们采用 ABTS 自由基清除活性测定法、铜离子还原抗氧化能力测定法、浊度法和 DNA 切口保护测定法评估了重组 AcTXNL1 蛋白(rAcTXNL1)的体外抗氧化能力。rAcTXNL1 在所有测定中均表现出浓度依赖性的强抗氧化活性。此外,在细胞环境中,过表达 AcTXNL1 可提高 HO 应激下的细胞活力,并降低脂多糖(LPS)诱导的一氧化氮(NO)产生。总之,实验结果表明 AcTXNL1 是双锯鱼中一种具有抗氧化和免疫重要性的基因。