Cytochrome and alternative respiratory pathways compete for electrons in the presence of pyruvate in soybean mitochondria.
The partitioning of electrons between the alternative oxidase and the cytochrome pathway of soybean mitochondria has been reassessed in the presence of the alternative oxidase activator pyruvate. In the presence of pyruvate and with succinate as substrate, the alternative oxidase became active at a much lower level of ubiquinone reduction than in the absence of pyruvate. Under state 4 (no ADP present) conditions, activation of the alternative oxidase with pyruvate resulted in an oxidation of b cytochromes, demonstrating switching of electrons away from the cytochrome chain. In the presence of ferricyanide and the cytochrome oxidase inhibitor KCN, cytochrome chain activity could be followed spectrophotometrically and that of the alternative pathway with an oxygen electrode. Under these conditions, the addition of pyruvate diverted electron flow from the cytochrome chain to the alternative pathway; subsequent inhibition of the alternative oxidase increased electron flow via the cytochrome chain. This indicates that electrons can be switched from one pathway to the other when the cytochrome chain is not saturated and this was confirmed by n-propylgallate titrations (p plots) of mitochondria oxidizing succinate. Decreases in ADP/O ratios and phosphorylation rate upon addition of pyruvate indicated that the alternative pathway could also contribute to respiration under state 3 conditions. The results indicate that when the alternative oxidase is activated by pyruvate, it can compete for electrons with the cytochrome chain and does not act as an overflow pathway. The significance of these observations for in vivo respiration is discussed.
Hoefnagel
M H
MH
Botany Department, University of Adelaide, Australia.
Millar
A H
AH
Wiskich
J T
JT
Day
D A
DA
D003580:Cytochromes / Q000378:metabolism*; D004579:Electron Transport / Q000187:drug effects; D007700:Kinetics; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism*; D024101:Mitochondrial Proteins; D010085:Oxidative Phosphorylation / Q000187:drug effects; D010088:Oxidoreductases / Q000378:metabolism; D010101:Oxygen Consumption / Q000187:drug effects; D010940:Plant Proteins; D011435:Propyl Gallate / Q000494:pharmacology; D011773:Pyruvates / Q000494:pharmacology*; D019289:Pyruvic Acid; D012639:Seeds / Q000378:metabolism; D013025:Glycine max / Q000378:metabolism*; D000099188:Alternative Oxidase
D003580:Cytochromes; D024101:Mitochondrial Proteins; D010088:Oxidoreductases; D010940:Plant Proteins; D011435:Propyl Gallate; D011773:Pyruvates; D019289:Pyruvic Acid; D000099188:Alternative Oxidase