Brain α-Ketoglutarate Dehydrogenase Complex: Kinetic Properties, Regional Distribution, and Effects of Inhibitors
Article first published online: 5 OCT 2006
Journal of Neurochemistry
Volume 47, Issue 5, pages 1376–1386, November 1986
How to Cite
Lai, J. C. K. and Cooper, A. J. L. (1986), Brain α-Ketoglutarate Dehydrogenase Complex: Kinetic Properties, Regional Distribution, and Effects of Inhibitors. Journal of Neurochemistry, 47: 1376–1386. doi: 10.1111/j.1471-4159.1986.tb00768.x
- Issue published online: 5 OCT 2006
- Article first published online: 5 OCT 2006
- Received December 19, 1985; accepted May 1, 1986.
- Brain α-ketoglutarate dehydrogenase complex;
- α-Keto acid dehydrogenase complex;
- Brain mitochondria;
- Neurotoxicity of ammonia;
- Hepatic encephalopathy;
- Calcium inhibition of dehydrogenases
Abstract The substrate and cofactor requirements and some kinetic properties of the α-ketoglutarate dehydrogenase complex (KGDHC; EC 22.214.171.124, EC 126.96.36.199, and EC 188.8.131.52) in purified rat brain mitochondria were studied. Brain mitochondrial KGDHC showed absolute requirement for α-ketoglutarate, CoA and NAD, and only partial requirement for added thiamine pyrophosphate, but no requirement for Mg2+ under the assay conditions employed in this study. The pH optimum was between 7.2 and 7.4, but, at pH values below 7.0 or above 7.8, KGDHC activity decreased markedly. KGDHC activity in various brain regions followed the rank order: cerebral cortex > cerebellum ≧ midbrain > striatum = hippocampus > hypothalamus > pons and medulla > olfactory bulb. Significant inhibition of brain mitochondrial KGDHC was noted at pathological concentrations of ammonia (0.2–2 mM). However, the purified bovine heart KGDHC and KGDHC activity in isolated rat heart mitochondria were much less sensitive to inhibition. At 5 mM both β-methylene-D,L-aspartate and D,L-vinylglycine (inhibitors of cerebral glucose oxidation) inhibited the purified heart but not the brain mitochondrial enzyme complex. At approximately 10 μM, calcium slightly stimulated (by 10–15%) the brain mitochondrial KGDHC.
At concentrations above 100 μM, calcium (IC50= 1 mM) inhibited both brain mitochondrial and purified heart KGDHC. The present results suggest that some of the kinetic properties of the rat brain mitochondrial KGDHC differ from those of the purified bovine heart and rat heart mitochondrial enzyme complexes. They also suggest that the inhibition of KGDHC by ammonia and the consequent effect on the citric acid cycle fluxes may be of pathophysiological and/or pathogenetic importance in hyperammonemia and in diseases (e.g., hepatic encephalopathy, inborn errors of urea metabolism, Reye's syndrome) where hyperammonemia is a consistent feature. Brain accumulation of calcium occurs in a number of pathological conditions. Therefore, it is possible that such a calcium accumulation may have a deleterious effect on KGDHC activity.