Purification of active recombinant trypanosome alternative oxidase

FEBS Letters

Fukai, Yoshihisa²  Kawai, Keisuke²  Nagai, Kazuo¹  Nihei, Coichi²  Suzuki, Takashi⁴  Ohta, Nobuo⁴  Kita, Kiyoshi²  Yabu, Yoshisada⁴  Minagawa, Nobuko³  Osanai, Arihiro² 

[1] Department of Applied Biological Chemistry, Chubu University, Kasugai, Aichi 487-8501, Japan;Department of Biomedical Chemistry, Graduate School of Medicine, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan;Department of Biochemistry, Niigata College of Pharmacy, Niigata 950-2081, Japan;Department of Molecular Parasitology, Nagoya City University, Graduate School of Medical Sciences, Nagoya 467-8601, Japan

关键词: Trypanosoma brucei brucei;    Trypanosome alternative oxidase;    Alternative oxidase;    Parasite mitochondria;    Ascofuranone;    Quinol oxidase;    AOX;    alternative oxidase;    rTAO;    recombinant trypanosome alternative oxidase;    TAO;    trypanosome alternative oxidase;   

DOI  :  10.1016/S0014-5793(03)00120-0

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 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.

【 授权许可】

Unknown