Purification of active recombinant trypanosome alternative oxidase
Purification of active recombinant trypanosome alternative oxidase
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DOI：
10.1016/s0014-5793(03)00120-0
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发表时间：
2003-03-13
期刊：
影响因子：
3.5
通讯作者：
Kita, K
中科院分区：
文献类型：
--
作者：
Nihei, C;Fukai, Y;Kita, K
锥虫交替氧化酶(TAO)是非洲锥虫细长血流形式呼吸链的末端氧化酶。TAO是一种细胞色素不依赖、氰化物不敏感的喹酚氧化酶。这些特征不同于细菌的喹酚氧化酶，后者是属于血红素-铜末端氧化酶超家族的蛋白质。无法提纯稳定的TAO严重阻碍了对替代氧化酶家族的生化研究。在本研究中，我们能够使用洗涤剂洋地黄素从大肠杆菌膜中纯化重组TAO，使其达到均一。对纯化的TAO的动力学分析表明，特异性抑制剂阿司可呋喃酮是泛喹酚氧化酶活性的竞争性抑制剂。(C)2003年，埃尔塞维尔科学公司代表欧洲生化学会联合会出版。
Trypanosome alternative oxidase (TAO) is the terminal oxidase of the respiratory chain in long slender bloodstream forms of African trypanosomes. TAO is a cytochrome-independent, cyanide-insensitive quinol oxidase. These characteristics are distinct from those of the bacterial quinol oxidases, proteins that belong to the heme-copper terminal oxidase superfamily. The inability to purify stable TAO has severely hampered biochemical studies of the alternative oxidase family. In the present study, we were able to purify recombinant TAO to homogeneity from Escherichia coli membranes using the detergent digitonin. Kinetic analysis of the purified TAO revealed that the specific inhibitor ascofuranone is a competitive inhibitor of ubiquinol oxidase activity. (C) 2003 Published by Elsevier Science B.V. on behalf of the Federation of European Biochemical Societies.