Thioredoxins (Trxs) are a family of small and highly conserved proteins which play crucial roles in the maintenance and regulation of the cellular redox homeostasis. In this study, the full-length cDNAs of thioredoxin 1 (TfTrx1) and thioredoxin-related protein of 14 kDa (TfTrp14) were isolated from roughskin sculpin (Trachidermus fasciatus). TfTrx1 is 662 bp in length with a 336-bp open reading frame (ORF) that encodes for a peptide with 111 amino acids, and TfTrp14 consists of 1066 bp with a 372-bp ORF that is translated to 123 amino acids. TfTrx1 and TfTrp14 contain highly conserved catalytic site motif CGPC and CPDC, respectively. Tissue distribution analysis indicated that both genes were broadly expressed in all examined tissues with the highest expression of TfTrx1 in the blood and TfTrp14 in the brain. In post-LPS and heavy metal challenge, the mRNA of both genes was significantly increased in the skin, liver, spleen, and brain at various times. The results of western blot detection displayed that the time of the induced maximum protein expression was 6-h post-LPS injection in the skin and liver, which were slightly delayed compared with that of 2 h at mRNA level. The recombinant TfTrp14 and TfTrx1 proteins were expressed in E. coli BL21 (DE3). The increase of the fluorescence intensity in rTfTrx1 and rTfTrp14 suggested the redox state changes in the microenvironment around tryptophan residues. Both of the recombinant proteins exhibited concentration-dependent disulfide reductase activity towards insulin, and the catalytic activity of rTfTrx1 was much higher than that of rTfTrp14.

硫氧还蛋白 (Trxs) 是一个小型且高度保守的蛋白质家族，在维持和调节细胞氧化还原稳态中起关键作用。在这项研究中，硫氧还蛋白 1 (TfTrx1) 和 14 kDa 硫氧还蛋白相关蛋白 (TfTrp14) 的全长 cDNA 是从粗皮鱼 ( Trachidermus fasciatus ) 中分离得到的。）。TfTrx1 长 662 bp，具有 336 bp 的开放阅读框 (ORF)，编码具有 111 个氨基酸的肽，TfTrp14 由 1066 bp 和 372 bp 的 ORF 组成，可翻译为 123 个氨基酸。TfTrx1 和 TfTrp14 分别包含高度保守的催化位点基序 CGPC 和 CPDC。组织分布分析表明，这两个基因在所有检查的组织中广泛表达，血液中 TfTrx1 和脑中 TfTrp14 的表达最高。在 LPS 和重金属攻击后，两种基因的 mRNA 在不同时间在皮肤、肝脏、脾脏和大脑中均显着增加。Western blot检测结果显示，在皮肤和肝脏注射LPS后6 h，诱导的最大蛋白表达时间比mRNA水平的2 h略有延迟。大肠杆菌BL21 (DE3)。rTfTrx1 和 rTfTrp14 中荧光强度的增加表明色氨酸残基周围微环境中的氧化还原状态发生了变化。两种重组蛋白对胰岛素均表现出浓度依赖性二硫键还原酶活性，且rTfTrx1的催化活性远高于rTfTrp14。