Purification, characterization, and amino acid sequence of thioredoxin from Corynebacterium nephridii.
Purification, characterization, and amino acid sequence of thioredoxin from Corynebacterium nephridii.
复制标题
肾棒杆菌硫氧还蛋白的纯化、表征和氨基酸序列。
DOI：
10.1016/s0021-9258(19)52524-8
复制
发表时间：
1981
期刊：
影响因子：
--
通讯作者：
H. Hogenkamp
中科院分区：
文献类型：
--
作者：
M. Meng;H. Hogenkamp
已使用常规技术从肾棒杆菌的无细胞提取物中将硫氧还蛋白纯化至同质。该硫氧还蛋白能够作为酵母甲硫氨酸亚砜还原酶、大肠杆菌硫氧还蛋白还原酶和莱希曼乳杆菌核糖核苷酸还原酶的还原底物。相反，在存在肾梭菌或大肠杆菌硫氧还蛋白还原酶的情况下，来自肾梭菌的硫氧还蛋白不能充当来自同一生物体的核糖核苷酸还原酶的还原底物。当用二硫苏糖醇还原时，来自 C. nephridii 的硫氧还蛋白能够激活菠菜叶绿体果糖-1, 6-二磷酸酶、NADP-苹果酸脱氢酶和磷酸核糖激酶。硫氧还蛋白的完整一级结构已通过自动 Edman 降解和羧肽酶 Y 消化完整蛋白质和金黄色葡萄球菌蛋白酶衍生的肽来确定。梭菌蛋白酶消化和溴化氰裂解。它由105个氨基酸残基组成。将肾小球藻硫氧还蛋白的氨基酸序列与大肠杆菌的硫氧还蛋白的氨基酸序列进行比较，揭示了这两种蛋白质之间惊人的同源性。蛋白质的氨基酸残基几乎有一半是相同的，同源区域包括硫氧还蛋白的活性位点和提供分子的两个结构域之间相互作用的区域。
A thioredoxin has been purified to homogeneity from cell free extract of Corynebacterium nephridii using conventional techniques. This thioredoxin is able to serve as the reducing substrate for yeast methionine sulfoxide reductase, for Escherichia coli thioredoxin reductase and for Lactobacillus leichmannii ribonucleotide reductase. In contrast thioredoxin from C. nephridii is unable to serve as a reducing substrate for ribonucleotide reductase from the same organism in the presence of either C. nephridii or E. coli thioredoxin reductase. When reduced with dithiothreitol thioredoxin from C. nephridii is able to activate spinach chloroplast fructose-1, 6-diphosphatase, NADP-malate dehydrogenase and phosphoribulokinase.The complete primary structure of the thioredoxin has been determined by automated Edman degradation and carboxypeptidase Y digestion of the intact protein and of peptides derived from Staphylococcus aureus protease and clostripain digestion and cyanogen bromide cleavage. It consists of 105 amino acid residues. Comparison of the amino acid sequence of C. nephridii thioredoxin with that of thioredoxin from E. coli reveals an amazing homology between these two proteins. Almost half of the amino acid residues of the proteins are identical, the regions of homology include the active site of thioredoxin and the regions which provide the interaction between the two domains of the molecule.