Redox state of quinone affects sensitivity of Acanthamoeba castellanii mitochondrial uncoupling protein to purine nucleotides.
We studied FFA (free fatty acid)-induced uncoupling activity in Acanthamoeba castellanii mitochondria in the non-phosphorylating state. Either succinate or external NADH was used as a respiratory substrate to determine the proton conductance curves and the relationships between respiratory rate and the quinone reduction level. Our determinations of the membranous quinone reduction level in non-phosphorylating mitochondria show that activation of UCP (uncoupling protein) activity leads to a PN (purine nucleotide)-sensitive decrease in the quinone redox state. The gradual decrease in the rate of quinone-reducing pathways (using titration of dehydrogenase activities) progressively leads to a full inhibitory effect of GDP on LA (linoleic acid) induced proton conductance. This inhibition cannot be attributed to changes in the membrane potential. Indeed, the lack of GDP inhibitory effect observed when the decrease in respiratory rate is accompanied by an increase in the quinone reduction level (using titration of the quinol-oxidizing pathway) proves that the inhibition by nucleotides can be revealed only for a low quinone redox state. It must be underlined that, in A. castellanii non-phosphorylating mitochondria, the transition of the inhibitory effect of GDP on LA-induced UCP-mediated uncoupling is observed for the same range of quinone reduction levels (between 50% and 40%) as that observed previously for phosphorylating conditions. This observation, drawn from the two different metabolic states of mitochondria, indicates that quinone could affect UCP activity through sensitivity to PNs.
Swida
Aleksandra
A
Laboratory of Bioenergetics, Faculty of Biology, Adam Mickiewicz University, Umultowska 89, 61-614 Poznan, Poland.
Woyda-Ploszczyca
Andrzej
A
Jarmuszkiewicz
Wieslawa
W
D048248:Acanthamoeba castellanii / Q000378:metabolism*; D000818:Animals; D016227:Benzoquinones / Q000737:chemistry*; D006153:Guanosine Diphosphate / Q000737:chemistry; D007473:Ion Channels / Q000737:chemistry*; D019787:Linoleic Acid / Q000737:chemistry; D008564:Membrane Potentials; D008928:Mitochondria / Q000378:metabolism; D024101:Mitochondrial Proteins / Q000737:chemistry*; D009243:NAD / Q000737:chemistry; D010084:Oxidation-Reduction; D010085:Oxidative Phosphorylation; D010100:Oxygen / Q000737:chemistry; D010766:Phosphorylation; D011522:Protons; D011685:Purine Nucleotides / Q000737:chemistry*; D000071256:Uncoupling Protein 1
D016227:Benzoquinones; D007473:Ion Channels; D024101:Mitochondrial Proteins; D011522:Protons; D011685:Purine Nucleotides; D000071256:Uncoupling Protein 1; D009243:NAD; D006153:Guanosine Diphosphate; C004532:quinone; D019787:Linoleic Acid; D010100:Oxygen