Purification and properties of the bovine liver mitochondrial dihydroorotate dehydrogenase.
Dihydroorotate dehydrogenase has been purified 6,000-fold from bovine liver mitochondria to apparent homogeneity in six steps. Electrophoretic migration of the homogeneous enzyme on sodium dodecyl sulfate-polyacrylamide gels reveals a subunit Mr of 42,000. By contrast to the well-characterized, cytosolic dihydroorotate oxidases (EC 1.3.3.1), the purified bovine dehydrogenase is a dihydroorotate:ubiquinone oxidoreductase. Maximal rates of orotate formation are obtained using coenzymes Q6 or Q7 as cosubstrate electron acceptors. Concomitant with substrate oxidation, the enzyme will reduce simple quinones, such as benzoquinone, but at significantly lower rates (10-15%) than that obtained for reduction of coenzyme Q6. Enzyme-catalyzed substrate oxidation is not supported by molecular oxygen. The specificity of the purified enzyme for dihydropyrimidine substrates has also been explored. The methyl-, ethyl-, t-butyl-, and benzyl-S-dihydroorotates are substrates, but 1- and 3-methyl and 1,3-dimethyl methyl-S-dihydroorotates are not. Competitive inhibitors include product orotate, 5-methyl orotate, and racemic cis-5-methyl dihydroorotate.
D000818:Animals; D002417:Cattle; D000090722:Dihydroorotate Dehydrogenase; D006863:Hydrogen-Ion Concentration; D008930:Mitochondria, Liver / Q000201:enzymology*; D008970:Molecular Weight; D009491:Neurospora / Q000201:enzymology; D010088:Oxidoreductases / Q000302:isolation & purification* / Q000378:metabolism; D044925:Oxidoreductases Acting on CH-CH Group Donors; D011809:Quinones / Q000378:metabolism; D051381:Rats; D013379:Substrate Specificity; D014451:Ubiquinone / Q000378:metabolism
D000090722:Dihydroorotate Dehydrogenase; D011809:Quinones; D014451:Ubiquinone; D010088:Oxidoreductases; D044925:Oxidoreductases Acting on CH-CH Group Donors