Gene cloning, sequence analysis, and expression of 2-methyl-3-hydroxypyridine-5-carboxylic acid oxygenase
Gene cloning, sequence analysis, and expression of 2-methyl-3-hydroxypyridine-5-carboxylic acid oxygenase
复制标题
DOI：
10.1073/pnas.94.14.7233
复制
发表时间：
1997-07-08
影响因子：
11.1
通讯作者：
Massey, V
中科院分区：
文献类型：
--
作者：
Chaiyen, PC;Ballou, DP;Massey, V
通过使用对应于该酶的N末端的寡核苷酸探针来克隆编码2-甲基-3-羟基吡啶-5-羧酸加氧酶(MHPCO；EC 1.14.12.4)的基因，以筛选假单胞菌属物种的DNA文库。 MA-1。该基因编码一个由 379 个氨基酸残基组成的蛋白质，对应的分子量为 41.7 kDa，与之前估计的 MHPCO 相同。 MHPCO 在大肠杆菌中表达，并被发现与假单胞菌的天然酶具有相同的特性。 MA-1。这项研究表明，MHPCO 是一种同源四聚体蛋白，每个亚基结合一个黄素腺嘌呤二核苷酸。该酶与其他羟化酶的序列比较揭示了芳香族黄素蛋白羟化酶中的保守区域。
The gene encoding 2-methyl-3-hydroxypyridine-5-carboxylic acid oxygenase (MHPCO; EC 1.14.12.4) was cloned by using an oligonucleotide probe corresponding to the N terminus of the enzyme to screen a DNA library of Pseudomonas sp. MA-1. The gene encodes for a protein of 379 amino acid residues corresponding to a molecular mass of 41.7 kDa, the same as that previously estimated for MHPCO. MHPCO was expressed in Escherichia coli and found to have the same properties as the native enzyme from Pseudomonas sp. MA-1. This study shows that MHPCO is a homotetrameric protein with one flavin adenine dinucleotide bound per subunit. Sequence comparison of the enzyme with other hydroxylases reveals regions that are conserved among aromatic flavoprotein hydroxylases.