Studies on the influence of fatty acids on pyruvate dehydrogenase interconversion in rat-liver mitochondria.

1. The effect of fatty acids on the interconversion of pyruvate dehydrogenase between its active (nonphosphorylated) and inactive (phosphorylated) forms was measured in rat liver mitochondria respiring in state 3 with pyruvate plus malate and 2-oxoglutarate plus malate and during state 4 to state 3 transition in the presence of different substrates. The content of intramitochondrial adenine nucleotides was determined in the parallel experiments. 2. Decrease of the intramitochondrial ATP/ADP ratio with propionate and its increase with palmitoyl-L-carnitine in state 3 is accompanied by a shift of the steady-state of the pyruvate dehydrogenase system towards the active or the inactive form, respectively. 3. Transition from the high energy state (state4) to the active respiration (state3) in mitochondria oxidizing 2-oxoglutarate or plamitoyl-L-carnitine causes an increase of the amount of the active form of pyruvate dehydrogenase due to the decrease of ATP/ADP ratio in the matrix. 4. No change in ATP/ADP ratio can be observed in the presence of octanoate in mitochondria oxidizing pyruvate or 2-oxoglutarate in state 3 or during state 4 to state 3 transition. Simultanelusly, no significant change in phosphorylation state of pyruvate dehydrogenase occurs and a low amount of the enzyme in the active form is present with octanoate or octanoate plus 2-oxoglutarate. Pyruvate abolishes this effect of octanoate and shifts the steady-state of pyruvate dehydrogenase system towards the active form. 5. These results indicate that fatty acids influence the interconversion of pyruvate dehydrogenase mainly by changing intramitochondrial ATP/ADP ratio. However, the comparison of the steady-state level of the pyruvate dehydrogenase system in the presence of different substrates in various metabolic conditions provides some evidence that accumulation of acetyl-CoA and high level of NADH may promote the phosphorylation of pyruvate dehydrogenase. 6. Pyruvate exerts its protective effect against phosphorylation of pyruvate dehydrogenase in the presence of fatty acids of short, medium or long chain in a manner which depends on its concentration. It is suggested that in isolated mitochondria pyruvate counteracts the effect of acetyl-CoA and NADH on pyruvate dehydrogenase kinase.