title: High membrane potential promotes alkenal-induced mitochondrial uncoupling and influences adenine nucleotide translocase conformation.
abstract: Mitochondria generate reactive oxygen species, whose downstream lipid peroxidation products, such as 4-hydroxynonenal, induce uncoupling of oxidative phosphorylation by increasing proton leak through mitochondrial inner membrane proteins such as the uncoupling proteins and adenine nucleotide translocase. Using mitochondria from rat liver, which lack uncoupling proteins, in the present study we show that energization (specifically, high membrane potential) is required for 4-hydroxynonenal to activate proton conductance mediated by adenine nucleotide translocase. Prolonging the time at high membrane potential promotes greater uncoupling. 4-Hydroxynonenal-induced uncoupling via adenine nucleotide translocase is prevented but not readily reversed by addition of carboxyatractylate, suggesting a permanent change (such as adduct formation) that renders the translocase leaky to protons. In contrast with the irreversibility of proton conductance, carboxyatractylate added after 4-hydroxynonenal still inhibits nucleotide translocation, implying that the proton conductance and nucleotide translocation pathways are different. We propose a model to relate adenine nucleotide translocase conformation to proton conductance in the presence or absence of 4-hydroxynonenal and/or carboxyatractylate.
authors.affiliation: MRC Dunn Human Nutrition Unit, Wellcome Trust/MRC Building, Hills Road, Cambridge, CB2 0XY, UK.
authors.forename: Vian
authors.initials: V
authors.lastname: Azzu
authors.forename: Nadeene
authors.initials: N
authors.lastname: Parker
authors.forename: Martin D
authors.initials: MD
authors.lastname: Brand
mesh_terms: D000447:Aldehydes / Q000737:chemistry; D000818:Animals; D001692:Biological Transport; D005260:Female; D007700:Kinetics; D008564:Membrane Potentials; D008928:Mitochondria / Q000378:metabolism*; D008930:Mitochondria, Liver / Q000378:metabolism; D000226:Mitochondrial ADP, ATP Translocases / Q000737:chemistry*; D009711:Nucleotides / Q000737:chemistry; D010085:Oxidative Phosphorylation; D010101:Oxygen Consumption; D011487:Protein Conformation; D011522:Protons; D051381:Rats
chemical_list: D000447:Aldehydes; D009711:Nucleotides; D011522:Protons; D000226:Mitochondrial ADP, ATP Translocases; C027576:4-hydroxy-2-nonenal