Functional properties of the Ustilago maydis alternative oxidase under oxidative stress conditions.
The mitochondrial respiratory chain of Ustilago maydis contains two terminal oxidases, the cytochrome c oxidase (COX) and the alternative oxidase (AOX). To understand the biochemical events that control AOX activity, we studied the regulation and function of AOX under oxidative stress. The activity of this enzyme was increased by both pyruvate (K(05)=2.6 mM) and purine nucleotides (AMP, K(05)=600 microM) in mitochondria using succinate as respiratory substrate. When U.maydis cells were grown in the presence of antimycin A, the amount of AOX in mitochondria was markedly increased and its selectivity towards AMP and pyruvate changed, suggesting that post-translational events may play a role in the regulation of AOX activity under stress conditions. Addition of antimycin A to isolated mitochondria induced the inactivation of AOX, the formation of lipid peroxides and the loss of glutathione from mitochondria. The two last processes are probably related with the time dependent inactivation of AOX, in agreement with the inhibition of the enzyme by tert-butyl hydroperoxide. Our results suggest that the in vivo operation of AOX in U. maydis depends on the mitochondrial antioxidant machinery, including the glutathione linked systems.
Sierra-Campos
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E
Departamento de Bioquímica, Facultad de Medicina, Universidad Nacional Autónoma de México, Apartado Postal 70-159, Coyoacán 04510, México DF, México.
Velázquez
I
I
Matuz-Mares
D
D
Villavicencio-Queijeiro
A
A
Pardo
J P
JP
D000968:Antimycin A / Q000494:pharmacology; D020536:Enzyme Activators / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005978:Glutathione / Q000378:metabolism; D008054:Lipid Peroxides / Q000378:metabolism; D008928:Mitochondria / Q000201:enzymology*; D024101:Mitochondrial Proteins; D018384:Oxidative Stress; D010088:Oxidoreductases / Q000378:metabolism*; D010940:Plant Proteins; D011687:Purines / Q000494:pharmacology; D019289:Pyruvic Acid / Q000494:pharmacology; D019802:Succinic Acid / Q000378:metabolism; D014588:Ustilago / Q000201:enzymology*; D000099188:Alternative Oxidase
D000968:Antimycin A; D020536:Enzyme Activators; D004791:Enzyme Inhibitors; D005978:Glutathione; D008054:Lipid Peroxides; D024101:Mitochondrial Proteins; D010088:Oxidoreductases; D010940:Plant Proteins; D011687:Purines; D019289:Pyruvic Acid; D019802:Succinic Acid; D000099188:Alternative Oxidase