Calcium and 2-oxoglutarate-mediated control of aspartate formation by rat heart mitochondria

Abstract

Studies of the influence of calcium on the metabolism of cardiac mitochondria have indicated that calcium activates key enzymes involved in the citric acid cycle. Calcium-mediated activation of one of these enzymes, 2-oxoglutarate dehydrogenase, has been shown to cause a marked decrease in the steady-state concentration of 2-oxoglutarate in both heart and liver mitochondria. In liver, 2-oxoglutarate is a potent inhibitor of oxalacetate transamination to aspartate and activation of this enzyme by calcium-mobilizing hormones leads to a stimulation of aspartate formation and gluconeogenesis. Since mitochondrial aspartate formation is a key step in the malate/aspartate shuttle, we investigated the control of aspartate formation by cardiac mitochondria. In mitochondria incubated with glutamate and malate, activation of 2-oxoglutarate dehydrogenase by calcium led to an inhibition of aspartate formation. However, calcium caused a stimulation of aspartate production when incubations were supplemented with pyruvate as an additional substrate. Estimates of the mitochondrial redox potential (NADH/NAD+) indicated that both calcium and pyruvate increased the redox potential. The observed influence of calcium on aspartate formation was found to be due to a balance between is inhibitory effect, caused by an increased redox potential, and its stimulatory effect, caused by a decreased 2-oxoglutarate concentration. Under conditions in which the redox component was held constant, a kinetic analysis indicated that the apparent Ki for 2-oxoglutarate inhibition of aspartate formation is 0.2 mM. The data suggest that activation of cardiac 2-oxoglutarate dehydrogenase by calcium could lead to stimulation of the mitochondrial oxidation of cytosolic NADH via the malate/aspartate cycle.