Support of respiration and citrulline synthesis by isolated hepatic mitochondria from Squalus acanthias by acyl-coas and other nitrogen-donating substrates

Abstract

1. 1. The effects of different substrates supporting respiration and glutamine-dependent citrulline synthesis from ornithine, ammonia, and bicarbonate by isolated hepatic mitochondria from Squalus acanthias (spiny dogfish) were determined. 2. 2. Highest rates of respiration were achieved with succinate, palmitoyl-CoA, and β-hydroxybutyrate as oxidizable substrates. All acyl-CoAs tested (from C-2 to C-22) supported carnitine-dependent respiration at a substantial rate. Short-chain fatty acids did not support respiration. 3. 3. Ammonia required for citrulline synthesis could be formed from glutamate, or from leucine plus α-ketoglutarate which gives rise to glutamate by transamination, as the result of glutamate dehydrogenase activity, but the reaction was inhibited by succinate or other oxidizable substrates. Alanine or ornithine could not be substituted for leucine, suggesting that leucine may specifically activate glutamate dehydrogenase. 4. 4. Glutamate required for citrulline synthesis could be formed from α-ketoglutarate and ammonia as the result of glutamate dehydrogenase activity if succinate was present. Transamination of α-ketoglutarate with ornithine present in the reaction mixtures provided glutamate at a rapid rate whether or not succinate was present. 5. 5. These results are consistent with the view that hepatic dogfish mitochondria efficiently utilize acyl-CoAs derived from triglyceride stores in the liver to support respiration, glutamine-dependent citrulline synthesis from ammonia, and formation of ketone bodies as a major fuel for muscle.