Phosphate-induced efflux of adenine nucleotides from rat-heart mitochondria: evaluation of the roles of the phosphate/hydroxyl exchanger and the dicarboxylate carrier.
Upon the addition of inorganic phosphate, isolated rat-heart mitochondria released endogenous adenine nucleotides. To elucidate the mechanism of this phosphate-induced efflux, we evaluated the relative roles of three inner mitochondrial membrane carriers: the adenine nucleotide translocase, the phosphate/hydroxyl exchanger, and the dicarboxylate carrier. Atractyloside (a specific inhibitor of the adenine nucleotide translocase) prevented this efflux, but did not inhibit mitochondrial swelling. Inhibitors of the phosphate/hydroxyl exchanger (200 microM n-ethylmaleimide and 10 microM mersalyl) did not inhibit phosphate-induced efflux. 200 microM mersalyl (which inhibited both the phosphate/hydroxyl exchanger and the dicarboxylate carrier) inhibited the rate of efflux approx. 65% Phenylsuccinate and 2-n-butylmalonate (inhibitors of the dicarboxylate carrier) partially inhibited phosphate-induced efflux and adenine nucleotide translocase activity. Mersalyl (200 microM) had no effect on adenine nucleotide translocase activity. Partial inhibition of the adenine nucleotide translocase by phenylsuccinate and butylmalonate could not explain the extent of inhibition of phosphate-efflux by these agents. Moreover, the rates of adenine nucleotide efflux in the presence of phenylsuccinate, butylmalonate, or mersalyl correlated well with the ability of these agents to inhibit succinate-supported respiration. We conclude that phosphate-induced efflux of adenine nucleotides from rat heart mitochondria occurs over the adenine nucleotide translocase, and that the site of action of the phosphate is not the phosphate/hydroxyl exchanger, but is likely the dicarboxylate carrier.
D000227:Adenine Nucleotides / Q000378:metabolism*; D000818:Animals; D001278:Atractyloside / Q000494:pharmacology; D002352:Carrier Proteins / Q000378:metabolism*; D029382:Dicarboxylic Acid Transporters; D005033:Ethylmaleimide / Q000494:pharmacology; D006878:Hydroxides / Q000378:metabolism; D007700:Kinetics; D008297:Male; D008314:Malonates / Q000494:pharmacology; D008634:Mersalyl / Q000494:pharmacology; D008929:Mitochondria, Heart / Q000187:drug effects / Q000378:metabolism*; D000226:Mitochondrial ADP, ATP Translocases / Q000037:antagonists & inhibitors; D010101:Oxygen Consumption / Q000187:drug effects; D010710:Phosphates / Q000494:pharmacology*; D051381:Rats; D011919:Rats, Inbred Strains; D013386:Succinates / Q000378:metabolism / Q000494:pharmacology; D019802:Succinic Acid
D000227:Adenine Nucleotides; D002352:Carrier Proteins; D029382:Dicarboxylic Acid Transporters; D006878:Hydroxides; D008314:Malonates; D010710:Phosphates; D013386:Succinates; D001278:Atractyloside; C006860:2-n-butylmalonate; D008634:Mersalyl; C017625:2-phenylsuccinate; D000226:Mitochondrial ADP, ATP Translocases; D019802:Succinic Acid; D005033:Ethylmaleimide