title: The effect of pH on the alternative oxidase activity in isolated Acanthamoeba castellanii mitochondria.
abstract: Mitochondria of Acanthamoeba castellanii possess a cyanide-resistant GMP-stimulated ubiquinol alternative oxidase in addition to the cytochrome pathway. In a previous work it has been observed that an interaction between the two ubiquinol-oxidizing pathways exists in intact A. castellanii mitochondria and that this interaction may be due to a high sensitivity of the alternative oxidase to matrix pH. In this study we have shown that the alternative oxidase activity reveals a pH-dependence with a pH optimum at 6.8 whatever the reducing substrate may be. The GMP stimulation of alternative oxidase is also strongly dependent on pH implicating probably protonation/deprotonation processes at the level of ligand and protein with an optimum pH at 6.8. The ubiquinone redox state-dependence of alternative oxidase activity is modified by pH in such a way that the highest activity for a given ubiquinone redox state is observed at pH 6.8. Thus pH, binding of GMP, and redox state of ubiquinone collaborate to set the activity of the GMP-stimulated alternative oxidase in isolated A. castellanii mitochondria. The high pH sensitivity of the alternative oxidase could link inactivation of the cytochrome pathway proton pumps to activation of the alternative oxidase with acceleration of redox free energy dissipation as a consequence.
authors.affiliation: Department of Bioenergetics, Institute of Molecular Biology and Biotechnology, Adam Mickiewicz University, Poznań, Poland.
authors.forename: Wiesława
authors.initials: W
authors.lastname: Jarmuszkiewicz
authors.forename: Lilla
authors.initials: L
authors.lastname: Hryniewiecka
authors.forename: Francis E
authors.initials: FE
authors.lastname: Sluse
mesh_terms: D000048:Acanthamoeba / Q000201:enzymology* / Q000648:ultrastructure; D000818:Animals; D019069:Cell Respiration; D004579:Electron Transport; D006157:Guanosine Monophosphate / Q000378:metabolism / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D008928:Mitochondria / Q000201:enzymology* / Q000378:metabolism; D024101:Mitochondrial Proteins; D010084:Oxidation-Reduction; D010088:Oxidoreductases / Q000378:metabolism*; D010940:Plant Proteins; D011190:Potassium Cyanide / Q000494:pharmacology; D014451:Ubiquinone / Q000378:metabolism; D000099188:Alternative Oxidase
chemical_list: D006157:Guanosine Monophosphate; D024101:Mitochondrial Proteins; D010088:Oxidoreductases; D010940:Plant Proteins; D011190:Potassium Cyanide; D014451:Ubiquinone; D000099188:Alternative Oxidase