Abstract
Pyrimidines and purine (deoxy)nucleotides are the building blocks of DNA and RNA. Nucleoside diphosphate sugars, e.g. UDP-glucose, are the reactive intermediates in the synthesis of nearly all glycosidic bonds between sugars.
In mammals the requirement for pyrimidines is met by UMP de novo synthesis and, to a greater or lesser extent, by salvage of free nucleosides. The exceptional compartmentation of the de novo synthesis with respect to mitochondrially-bound dihydroorotate dehydrogenase (’DHOdehase’ or ‘DHODH’, EC 1.3.99.11) is one focus of the present work. DHODH activity was determined by the dihydroorotate-dependent oxygen consumption or by the UV absorption of the product orotate with mitochondria isolated from rodent and porcine tissues. For comparison, the cytochrome c and choline-dependent oxygen consumption of mitochondria from different tissues was measured. The highest specific activity of the rat DHODH was found in liver (2.3 × 10-3 μmol/min × mg protein) > kidney > heart. The application of known enzyme inhibitors Brequinar Sodium and Leflunomide for DHODH and sodium cyanide for cytochrome c oxidase verified the specificity of the activity tests used. The relation of DHODH activity versus that of cytochrome c oxidase revealed the lowest ratios in heart mitochondria and the highest in liver mitochondria. Since disorders in the mitochondrial energy metabolism could entail severe impairment of pyrimidine biosynthesis via respiratory-chain coupled DHODH, it is suggested to include improvement of pyrimidine nucleotide status in therapy protocols. (Mol Cell Biochem 174: 115–119, 1997)
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References
Jones ME: Pyrimidine nucleotide biosynthesis in animals. Genes, enzymes and regulation of UMP biosynthesis. Ann Rev Biochem 49: 253–279, 1980
Nagy M, Lacroute F: Divergent evolution of pyrimidine biosynthesis between anaerobic and aerobic yeast. Proc Nat Acad Sci USA 89: 8966–8970, 1992
Löffler M: A cytokinetic approach to determine the range of oxygen dependence of pyrimidine(deoxy)nucleotide biosynthesis relevant for cell proliferation. Cell Prolif 25: 169–179, 1992
Lakaschus G, Löffler M: Differential susceptibility of dihydroorotate dehydrogenase/oxidase to Brequinar Sodium (NSC 368390) in vitro. Biochem Pharmacol 43: 1025–1030, 1992
Knecht W, Bergjohann U, Gonski S, Kirschbaum B, Löffler M: Functional expression of a human dihydroorotate dehydrogenase fragment using the baculovirus expression vector system and kinetic investigation of the purified recombinant enzyme. Eur J Biochem 240: 6292–6301, 1996
Angermüller S, Löffler M: Location of dihydroorotate oxidase in myocardium and kidney cortex of the rat. An electron microscopic study using the cerium technique. Histochem Cell Biol 103: 287–292, 1995
öffler M, Becker C, Wegerle E, Schuster G: Catalytic enzyme histochemistry and biochemical analysis of dihydroorotate dehydrogenase/oxidase and succinate dehydrogenase in mammalian tissues, cells and mitochondria. Histochem Cell Biol 105: 119–128
Mela L, Seitz S: Isolation of mitochondria with emphasis on heart mitochondria from small amount of tissue. Meth Enzymol 55: 39–44, 1979
Darley-Usmar VM, Rickwood D, Wilson MT: Mitochondria, A Practical Approach, IRL Press, Oxford, 1987
Greene S, Watanabe K, Braatscz-Trulson J, Lou L: Inhibition of dihydroorotate dehydrogenase by the immunosuppressive agent leflunomide. Biochem Pharmacol 50: 861–867, 1995
Taylor RW: The control of mitochondrial oxidations by complex III in rat muscle and liver mitochondria. J Biol Chem 269: 3523–3528, 1994
Haubrich DR, Gerber NH: Choline dehydrogenase. Biochem Pharmacol 30:2993–3000, 1981
Miller B, Schmid H, Schmolke M, Guder WG: Determination of choline dehydrogenase activity along the rat nephron. Biol Chem Hoppe Seyler 377: 129–137, 1996
Suttle DP, Becroft DMO, Webster DR: Hereditary orotic aciduria and other disorders of pyrimidine metabolism. In: CR Scriver, AL Beaudet, WS Sly, D Valle (eds). The Metabolic Basis of Inherited Disease. Mc Graw-Hill, London, pp 1095–1126, 1989
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Löffler, M., Jöckel, J., Schuster, G., Becker, C. (1997). Dihydroorotat-ubiquinone oxidoreductase links mitochondria in the biosynthesis of pyrimidine nucleotides. In: Gellerich, F.N., Zierz, S. (eds) Detection of Mitochondrial Diseases. Developments in Molecular and Cellular Biochemistry, vol 21. Springer, Boston, MA. https://doi.org/10.1007/978-1-4615-6111-8_19
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DOI: https://doi.org/10.1007/978-1-4615-6111-8_19
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