细胞内抗氧化剂谷氧还蛋白控制细胞增殖和存活。基于活跃的网站,结构,和保守的域基序,它分为两类。I类含有在共有活性位点序列CXXC中具有两个半胱氨酸的二硫醇Grxs,而II类具有在活性位点具有一个半胱氨酸残基的单硫醇Grxs。单硫醇Grxs还可以具有另外的N-末端硫氧还蛋白(Trx)样结构域。以前,我们报道了来自九头蛇(HvGrx1)的Grx1的表征,这是一种二硫醇同工型。这里,我们报道了分子克隆,表达式,分析,以及Grx的另一种同工型的表征,这是来自Hydravulgaris(HvGrx3)的多结构域单硫醇谷氧还蛋白-3。它编码具有303个氨基酸的蛋白质,并且比HvGrx1明显更大,并且差异更大。计算机分析表明,Grx1和Grx3具有22.5%和9.9%的相同核苷酸和氨基酸序列,分别。HvGrx3在其氨基末端有两个谷氧还蛋白结构域和一个硫氧还蛋白样结构域,与HvGrx1不同,HvGrx1具有单个谷氧还蛋白结构域。像其他单硫醇谷氧还蛋白一样,HvGrx3未能还原谷胱甘肽-羟乙基二硫化物。在整个九头蛇,发现HvGrx3在整个身体柱上都有表达,并且用H2O2处理导致HvGrx3的显著上调。当转染HCT116(人结肠癌细胞)细胞时,HvGrx3增强细胞增殖和迁移,表明这种同工型可能参与这些细胞功能。这些转染的细胞也更好地耐受氧化应激。
Intracellular antioxidant glutaredoxin controls cell proliferation and survival. Based on the active site, structure, and conserved domain motifs, it is classified into two classes. Class I contains dithiol Grxs with two cysteines in the consensus active site sequence CXXC, while class II has monothiol Grxs with one cysteine residue in the active site. Monothiol Grxs can also have an additional N-terminal thioredoxin (Trx)-like domain. Previously, we reported the characterization of Grx1 from
Hydra vulgaris (HvGrx1), which is a dithiol isoform. Here, we report the molecular cloning, expression, analysis, and characterization of another isoform of Grx, which is the multidomain monothiol glutaredoxin-3 from
Hydra vulgaris (HvGrx3). It encodes a protein with 303 amino acids and is significantly larger and more divergent than HvGrx1. In-silico analysis revealed that Grx1 and Grx3 have 22.5% and 9.9% identical nucleotide and amino acid sequences, respectively. HvGrx3 has two glutaredoxin domains and a thioredoxin-like domain at its amino terminus, unlike HvGrx1, which has a single glutaredoxin domain. Like other monothiol glutaredoxins, HvGrx3 failed to reduce glutathione-hydroxyethyl disulfide. In the whole
Hydra, HvGrx3 was found to be expressed all over the body column, and treatment with H2O2 led to a significant upregulation of HvGrx3. When transfected in HCT116 (human colon cancer cells) cells, HvGrx3 enhanced cell proliferation and migration, indicating that this isoform could be involved in these cellular functions. These transfected cells also tolerate oxidative stress better.