Bicarbonate and the pathway of glutamate oxidation in isolated rat-liver mitochondria.

1. The factors affecting the pathway of glutamate oxidation were studied in isolated rat-liver mitochondria in incubations of 2-3 min. 2. It was found that bicarbonate at a physiological concentration has a profound effect on the pathway of glutamate oxidation. Ammonia formation via glutamate dehydrogenase is stimulated by bicarbonate [from 5.48 +/- 0.29 (n = 10) to 9.57 +/- 0.73 (n = 8) nmol X min-1 X mg protein-1], whereas aspartate formation via the transamination pathway is inhibited [from 38.41 +/- 2.24 (n = 9) to 24.56 +/- 3.28 (n = 6) nmol X min-1 X mg protein-1]. 3. Bicarbonate has no effect on the rate of transport of glutamate via the glutamate-hydroxyl translocator. 4. The interaction of bicarbonate with the pathway of glutamate oxidation occurs primarily at the level of succinate dehydrogenase, due to competitive inhibition of the enzyme by bicarbonate. 5. Inhibition by bicarbonate of the transamination pathway leads to a decrease in intramitochondrial 2-oxoglutarate, so that the deamination pathway is stimulated. 6. Using an equation which describes flux through glutamate dehydrogenase kinetically, it could be shown that the bicarbonate-induced decrease in intramitochondrial 2-oxoglutarate quantitatively accounts for the enhanced rate of deamination. 7. It is concluded that in the intact liver flux through glutamate dehydrogenase is sufficient to account for the ammonia formation required for urea synthesis from substrates such as alanine.