Purification of active recombinant trypanosome alternative oxidase

被引：44

作者：

Nihei, C

Fukai, Y

Kawai, K

Osanai, A

Yabu, Y

Suzuki, T

Ohta, N

Minagawa, N

Nagai, K

Kita, K

机构：

[1] Univ Tokyo, Grad Sch Med, Dept Biomed Chem, Bunkyo Ku, Tokyo 1130033, Japan

[2] Nagoya City Univ, Grad Sch Med Sci, Dept Mol Parasitol, Nagoya, Aichi 4678601, Japan

[3] Niigata Coll Pharm, Dept Biochem, Niigata 9502181, Japan

[4] Chubu Univ, Dept Appl Biol Chem, Kasugai, Aichi 4878501, Japan

来源：

FEBS LETTERS | 2003年 / 538卷 / 1-3期

关键词：

Trypanosoma brucei brucei; Trypanosome alternative oxidase; alternative oxidase; parasite mitochondria; ascofuranone; quinol oxidase;

D O I：

10.1016/S0014-5793(03)00120-0

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

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.

引用

页码：35 / 40

页数：6

共 28 条

[1]

AFFEURTIT C, 2001, BIOCHIM BIOPHYS ACTA, V1504, P58

[2] Exploring the molecular nature of alternative oxidase regulation and catalysis [J].