The effect of temperature and chronic ethanol feeding on the proton electrochemical potential and phosphate potential in rat liver mitochondria.

The relationship between the proton electrochemical potential (delta mu H) and the maximal free energy of ATP hydrolysis (delta GP) in coupled respiring rat liver mitochondria was investigated as a function of temperature and chronic ethanol-feeding. The flow dialysis method was utilized to measure the temperature dependence of delta mu H from the uptake of 86Rb (in the presence of valinomycin) and [14C]DMO. delta GP in state 4 was determined by a null-point titration of the reversible, H+-coupled ATPase against the phosphate potential. delta mu H increases with temperature from 196 mV at 10 degrees C, to 217 mV at 40 degrees C. The maximal delta GP at state 4 decreases as a function of temperature from 67.8 kJ/mol at 10 degrees C, to 54.8 kJ/mol at 40 degrees C. As a result, the ratio delta GP/delta mu H decreases with temperature from 3.56 at 10 degrees C to 2.60 at 40 degrees C. Similar studies with mitochondria from rats which were chronically fed with ethanol show that, while delta GP at state 4 decreases in these rats from 61.2 to 56.0 (25 degrees C), the delta mu H is essentially unchanged at 212 mV. Thus the ratio delta GP/delta mu H in ethanol-fed rats at 25 degrees C is 2.77 as compared with 2.97 in control. Similar reduction of delta GP was observed in inverted inner membranes from ethanol-fed rats. Both the temperature dependence of delta GP/delta mu H and the effect of ethanol-feeding cannot be easily explained by the chemiosmotic hypothesis which postulates that delta mu H is the only driving force for ATP synthesis. In contrast, a parallel coupling model, which postulates that intramembrane proton transfer from redox pumps to ATPase is mediated by the formation of dynamic aggregates of the mitochondrial inner-membrane proteins, can easily accommodate these findings. Accordingly, the temperature effect is due to weakening of these fragile aggregates, while the ethanol-feeding effect is the result of reduced concentration of active pumps, which decrease the frequency of formation of functional aggregates.