Mitochondrial transmembrane pH and electrical gradients: evaluation of their energy relationships with respiratory rate and adenosine 5'-triphosphate synthesis.

Mitochondria from rat liver were suspended in ionic media, and parallel measurements were made of the respiratory rate, intramitochondrial volumes, and the phosphorylation state [[ATP]/([ADP][Pi])], as well as transmembrane pH gradients (from the distribution of labeled weak acids and bases) and electrical gradients (from the distribution of labeled lipid-soluble ions and of K+ in the presence of valinomycin). A decrease in the pH gradient was observed with additions of sodium propionate, acetate, or lactate. In the case of propionate, the pH gradient decreased from -0.94 +/- 0.10 (control) to -0.09 +/- 0.08 unit (with 3 mM propionate) without change in any of the other measured parameters. Variation of the intramitochondrial volumes was achieved by changing [K+] (+valinomycin) and by diluting the choline chloride media. The transmembrane electrical potential decreased from -150 mV under control conditions to approximately -75 mV in a 70 mosM medium or when 8 mM K+ (+valinomycin) was present. The changes in [ATP]/([ADP][Pi]) were as follows: from 1.8 X 10(4) (control) to 3.9 X 10(2) M-1 with 8 mM K+ (+valinomycin) or 1.0 X 10(4) M-1 when the osmolarity was decreased from 300 to 70 mosM. This corresponds to a decrease in the energy utilized for ATP synthesis from 14.1 to 11.9 kcal/mol (from 58.9 to 49.7 kJ/mol) with added K+ and to 13.8 kcal/mol (57.7 kJ/mol) with decreased osmolarity. No direct correlation was found between the changes in transmembrane electrical potential, pH gradient, or total proton electrochemical gradient and either the respiratory rate or [ATP]/([ADP][Pi]).