Rozell B, Bárcena J A, Martínez-Galisteo E, Padilla C A, Holmgren A
Department of Oral Pathology, Karolinska Institutet, Stockholm/Sweden.
Eur J Cell Biol. 1993 Dec;62(2):314-23.
Glutaredoxin catalyzes glutathione-dependent disulfide oxidoreduction reactions in a coupled system with NADPH, GSH and glutathione reductase and has an active site disulfide/dithiol with the sequence -Cys-Pro-Tyr-Cys-. Calf thymus glutaredoxin (thioltransferase), which contains two additional structural half-cystine residues, was purified to homogeneity, using a modification of the previously described isolation procedure. This method involved a pI-shift of glutaredoxin, obtained after oxidation of the fully reduced form with hydroxyethyl-disulfide, followed by CM-Sepharose chromatography. On both SDS- and IEF-gels the protein migrated as one band (M(r) 12,000). The pure protein was used to affinity-purify rabbit antiglutaredoxin antibodies obtained by immunization with the oxidized form of glutaredoxin. Using these antibodies the distribution of glutaredoxin was mapped in calf organs and tissues by Western blots and by immunohistochemistry. Glutaredoxin was demonstrated in all organs investigated. Western blots showed the presence of weak additional high molecular weight bands of unknown identity in certain organs. The immunohistochemical analyses revealed that glutaredoxin is highly expressed in a wide variety of cell types, both epithelial and mesenchymal. The distribution and occurrence in the calf organs was similar to that previously described for thioredoxin in the rat. There were some exceptions: e.g., follicular cells in the ovary did not contain immunohistochemically demonstrable glutaredoxin but expressed thioredoxin. Particularly striking were observations of strong glutaredoxin immunoreactivity in oocytes in the ovary and the pattern of glutaredoxin in epithelial tissue of the skin and tongue reflecting differential expression during cell differentiation. The distribution demonstrated that glutaredoxin serves functions apart from the originally described role as hydrogen donor for ribonucleotide reductase which only occurs in replicating cells. Such functions should relate particularly to glutathione-catalyzed protein disulfide oxidoreductions and cellular signalling by redox regulating mechanisms.
谷氧还蛋白在与NADPH、谷胱甘肽（GSH）和谷胱甘肽还原酶组成的偶联系统中催化依赖谷胱甘肽的二硫键氧化还原反应，其活性位点二硫键/二硫醇的序列为-Cys-Pro-Tyr-Cys-。小牛胸腺谷氧还蛋白（硫醇转移酶）含有另外两个结构半胱氨酸残基，通过对先前描述的分离程序进行改进，将其纯化至同质。该方法包括用羟乙基二硫将完全还原形式氧化后得到的谷氧还蛋白的pI位移，随后进行CM-琼脂糖凝胶色谱。在SDS凝胶和IEF凝胶上，该蛋白均迁移为一条带（相对分子质量12,000）。纯蛋白用于亲和纯化通过用氧化形式的谷氧还蛋白免疫获得的兔抗谷氧还蛋白抗体。使用这些抗体，通过蛋白质免疫印迹法和免疫组织化学法绘制了小牛器官和组织中谷氧还蛋白的分布图。在所研究的所有器官中均证实存在谷氧还蛋白。蛋白质免疫印迹显示，在某些器官中存在身份不明的较弱的额外高分子量条带。免疫组织化学分析表明，谷氧还蛋白在多种细胞类型中高表达，包括上皮细胞和间充质细胞。小牛器官中的分布和存在情况与先前在大鼠中描述的硫氧还蛋白相似。也有一些例外情况：例如，卵巢中的卵泡细胞不含免疫组织化学可证实的谷氧还蛋白，但表达硫氧还蛋白。特别引人注目的是观察到卵巢卵母细胞中强烈的谷氧还蛋白免疫反应性，以及皮肤和舌上皮组织中谷氧还蛋白的模式，反映了细胞分化过程中的差异表达。分布情况表明，谷氧还蛋白除了最初描述的作为核糖核苷酸还原酶的氢供体（仅发生在复制细胞中）的作用外，还具有其他功能。这些功能尤其应与谷胱甘肽催化的蛋白质二硫键氧化还原反应以及通过氧化还原调节机制进行的细胞信号传导有关。
