Overproduction of highly active trypanosome alternative oxidase in Escherichia coli heme-deficient mutant.
Cyanide-insensitive trypanosome alternative oxidase (TAO) is the terminal oxidase of the respiratory chain of long slender bloodstream forms of the African trypanosome, which causes sleeping sickness in humans and nagana in cattle. TAO has been targeted for the development of anti-trypanosomal drugs, because it does not exist in the host. In this study, we established a system for overproduction of highly active TAO in Eschericia coli heme-deficient mutant. Kinetic analysis of recombinant enzyme and TAO in Trypanosoma brucei brucei mitochondria revealed that recombinant TAO retains the properties of native enzyme, indicating that recombinant TAO is quite valuable for further biochemical study of TAO.
Fukai
Yoshihisa
Y
Department of Biomedical Chemistry, Graduate School of Medicine, University of Tokyo 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
Nihei
Coichi
C
Kawai
Keisuke
K
Yabu
Yoshisada
Y
Suzuki
Takasi
T
Ohta
Nobuo
N
Minagawa
Nobuko
N
Nagai
Kazuo
K
Kita
Kiyoshi
K
D000818:Animals; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D006418:Heme / Q000172:deficiency*; D008928:Mitochondria / Q000201:enzymology; D024101:Mitochondrial Proteins; D009154:Mutation; D010088:Oxidoreductases / Q000235:genetics* / Q000378:metabolism*; D010940:Plant Proteins; D012717:Sesquiterpenes / Q000494:pharmacology; D014346:Trypanosoma brucei brucei / Q000201:enzymology* / Q000235:genetics; D000099188:Alternative Oxidase
D006418:Heme; D024101:Mitochondrial Proteins; D010088:Oxidoreductases; D010940:Plant Proteins; D012717:Sesquiterpenes; D000099188:Alternative Oxidase; C006640:ascofuranone