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1

The contribution of uncoupling protein and ATP synthase to state 3 respiration in Acanthamoeba castellanii mitochondria

100%
Jarmuszkiewicz W., Czarna M., Sluse-Goffart C., Sluse F. E.
Acta Biochimica Polonica
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2004
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tom 51
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nr 2
Mitochondria of the amoeba Acanthamoeba castellanii possess a free fatty acid-ac­tivated uncoupling protein (AcUCP) that mediates proton re-uptake driven by the mi­tochondrial proton electrochemical gradient. We show that AcUCP activity diverts energy from ATP synthesis during state 3 mitochondrial respiration in a fatty acid-dependent way. The efficiency of AcUCP in mitochondrial uncoupling increases when the state 3 respiratory rate decreases as the AcUCP contribution is constant at a given linoleic acid concentration while the ATP synthase contribution decreases with respiratory rate. Respiration sustained by this energy-dissipating process remains constant at a given linoleic acid concentration until more than 60% inhibition of state 3 respiration by n-butyl malonate is achieved. The present study supports the validity of the ADP/O method to determine the actual contributions of AcUCP (activated with various linoleic acid concentrations) and ATP synthase in state 3 respiration of A. castellanii mitochondria fully depleted of free fatty acid-activated and describes how the two contributions vary when the rate of succinate dehydrogenase is decreased by succinate uptake limitation.
2

Effect of growth at low temperature on the alternative pathway respiration in Acanthamoeba castellanii mitochondria

80%
Jarmuszkiewicz W., Fraczyk O., Hryniewiecka L.
Acta Biochimica Polonica
|
2001
|
tom 48
|
nr 3
Mitochondria of amoeba Acanthamoeba castellanii in addition to the conventional cytochrome pathway possess, like plant mitochondria, a cyanide-resistant alternative quinol oxidase. In mitochondria isolated from amoeba batch culture grown tempo­rarily at low temperature (6°C), higher respiration was accompanied by lower cou­pling parameters as compared to control culture (grown at 28°C). In the presence of benzohydroxamate, respiratory rates and coupling parameters were similar in both types of mitochondria indicating that growth in cold conditions did not disturb the cytochrome pathway. Increased contribution of alternative oxidase in total mitochon­drial respiration in low-temperature-grown amoeba cells was confirmed by calcula­tion of its contribution using ADP/O measurements. Furthermore, in mitochondria from low-temperature- grown cells the content of the alternative oxidase was in­creased and correlated with the increase in the unstimulated and GMP-stimulated cya­nide-resistant respiratory activity. A possible physiological role of higher activity of alternative oxidase as response to growth at a low temperature in unicellular organ­isms, such as amoeba, is discussed.
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