Citrate effects on the Ca2+-loading capacity of isolated rat liver mitochondria: interaction of citrate and ATP.
The maximal amounts of Ca2+ being accumulated (delta Ca2+max) and H+ emitted (delta H+max) by Ca2+-loading mitochondria, with succinate (+rotenone) as respiratory substrate, were evaluated. delta Ca2+max was increased by providing either citrate or ATP to a Pi- and Mg2+-free medium. With citrate, delta H+max was only scarcely increased, so that the effect of the proton-carrying anion resulted essentially from an increase in the Ca2+/H+ ratio, i.e., from preservation of membrane potential. With ATP (+/- oligomycin), the Ca2+/H+ ratio was unaltered; i.e., the increase of delta Ca2+max was paralleled by a related increase in delta H+max. Mitochondria appeared to retain Ca at higher delta pH, i.e., at lower membrane potential, in the presence of ATP. With citrate and ATP together, both the Ca2+/H+ ratio and delta H+max were largely increased, and the product of these two terms, delta Ca2+max, was considerably enlarged. The effect of either citrate or ATP was markedly reinforced in the presence of the other anion. In addition to increasing the Ca2+/H+ ratio, citrate contributed to increasing delta H+max in the presence of ATP, i.e., apparently sensitized mitochondria to the action of ATP. A citrate-induced depression of Ca2+ cycling across the inner membrane, even though pronounced, did not account for the sensitization. Supraadditive effects of citrate and ATP persisted in the presence of MgCl2 and Pi, under conditions of massive Ca2+ loading, and may contribute to the high capacity of mitochondria, in situ, to retain calcium.
D000255:Adenosine Triphosphate / Q000494:pharmacology*; D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002951:Citrates / Q000494:pharmacology*; D004357:Drug Synergism; D066298:In Vitro Techniques; D008274:Magnesium / Q000494:pharmacology; D015636:Magnesium Chloride; D008930:Mitochondria, Liver / Q000187:drug effects / Q000378:metabolism*; D011199:Potentiometry; D011522:Protons; D051381:Rats
D002951:Citrates; D011522:Protons; D015636:Magnesium Chloride; D000255:Adenosine Triphosphate; D008274:Magnesium; D002118:Calcium