Substrate kinetics of the Acanthamoeba castellanii alternative oxidase and the effects of GMP.
In Acanthamoeba castellanii mitochondria, the apparent affinity values of alternative oxidase for oxygen were much lower than those for cytochrome c oxidase. For unstimulated alternative oxidase, the K(Mox) values were around 4-5 microM both in mitochondria oxidizing 1 mM external NADH or 10 mM succinate. For alternative oxidase fully stimulated by 1 mM GMP, the KK(Mox) values were markedly different when compared to those in the absence of GMP and they varied when different respiratory substrates were oxidized (K(Mox) was around 1.2 microM for succinate and around 11 microM for NADH). Thus, with succinate as a reducing substrate, the activation of alternative oxidase (with GMP) resulted in the oxidation of the ubiquinone pool, and a corresponding decrease in K(Mox). However, when external NADH was oxidized, the ubiquinone pool was further reduced (albeit slightly) with alternative oxidase activation, and the K(Mox) increased dramatically. Thus, the apparent affinity of alternative oxidase for oxygen decreased when the ubiquinone reduction level increased either by changing the activator or the respiratory substrate availability.
Jarmuszkiewicz
Wieslawa
W
Laboratory of Bioenergetics, Adam Mickiewicz University, Fredry 10, 61-701 Poznan, Poland. wiesiaj@amu.edu.pl
Czarna
Malgorzata
M
Sluse
Francis E
FE
D048248:Acanthamoeba castellanii / Q000201:enzymology* / Q000648:ultrastructure; D003580:Cytochromes / Q000378:metabolism; D004579:Electron Transport / Q000187:drug effects / Q000502:physiology; D006157:Guanosine Monophosphate / Q000494:pharmacology*; D007700:Kinetics; D008928:Mitochondria / Q000201:enzymology; D024101:Mitochondrial Proteins; D009243:NAD / Q000378:metabolism; D010084:Oxidation-Reduction; D010088:Oxidoreductases / Q000378:metabolism*; D010940:Plant Proteins; D013268:Stimulation, Chemical; D013379:Substrate Specificity; D019802:Succinic Acid / Q000378:metabolism; D014451:Ubiquinone / Q000378:metabolism; D000099188:Alternative Oxidase
D003580:Cytochromes; D006157:Guanosine Monophosphate; D024101:Mitochondrial Proteins; D009243:NAD; D010088:Oxidoreductases; D010940:Plant Proteins; D019802:Succinic Acid; D014451:Ubiquinone; D000099188:Alternative Oxidase