Involvement of cytochrome c oxidase subunit III in energy coupling.
The role of the conserved acidic residues of subunit III of cytochrome c oxidase (COIII) in energy transduction was investigated. Using a COIII deletion mutant of Paracoccus denitrificans, complemented with a plasmid expressing either the wild type (wt) COIII gene or site-directed mutants of the COIII gene, we measured cytochrome c oxidase-dependent ATP synthesis, respiration, and membrane potential. Cytochrome c oxidase-dependent ATP synthesis was attenuated in nonacidic mutants of either Glu98 (E98A and E98Q), or Asp259 (D259A) but not in the acidic mutant E98D. The rates of respiration in the energy conversion-defective mutants were as high as or higher than that in the wt. The cytochrome c oxidase-induced increment of membrane potential in the nonacidic mutants was similar to or higher than that in the wt. In contrast, when succinate-driven ATP synthesis was mediated solely by ubiquinol oxidase (e.g., in the presence of myxothiazol), the rates of ATP synthesis in the nonacidic mutants were higher than that in the wt. Moreover, myxothiazol, which inhibited succinate respiration as well as ATP synthesis in wt and E98D, stimulated ATP synthesis, while inhibiting succinate respiration, in the nonacidic mutants. These results indicate that the attenuation of energy conversion in these mutants is limited to cytochrome c oxidase and thus suggest that subunit III plays a role in energy conversion by cytochrome c oxidase.
Wu
S
S
Department of Pathology, Medical College of Pennsylvania, Philadelphia 19102, USA.
Moreno-Sanchez
R
R
Rottenberg
H
H
D000255:Adenosine Triphosphate / Q000032:analysis; D001205:Ascorbic Acid / Q000378:metabolism; D003576:Electron Transport Complex IV / Q000235:genetics / Q000378:metabolism*; D008564:Membrane Potentials; D008689:Methacrylates; D009154:Mutation; D010085:Oxidative Phosphorylation; D010101:Oxygen Consumption / Q000187:drug effects; D010230:Paracoccus / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*; D013386:Succinates / Q000378:metabolism; D019802:Succinic Acid; D013772:Tetramethylphenylenediamine / Q000378:metabolism; D013844:Thiazoles / Q000494:pharmacology; D014451:Ubiquinone / Q000031:analogs & derivatives / Q000378:metabolism
D008689:Methacrylates; D013386:Succinates; D013844:Thiazoles; D014451:Ubiquinone; C030517:myxothiazol; D000255:Adenosine Triphosphate; D019802:Succinic Acid; D003576:Electron Transport Complex IV; C003741:ubiquinol; D013772:Tetramethylphenylenediamine; D001205:Ascorbic Acid