Effect of 6-azauridine on de novo pyrimidine biosynthesis in cultured Ehrlich ascites cells. Orotate inhibition of dihydroorotase and dihydroorotate dehydrogenase.
The inhibition of dihydro-orotase (E 3.5.2.3) and dihydroorotate (DHO) dehydrogenase (dihydro-orotate oxidase, EC 1.3.3.1) by cellular orotate (OA) in Ehrlich ascites cells was studied by measuring the accumulation of the intermediates of de novo pyrimidine biosynthesis at various times after the addition of 6-azauridine to the culture medium. The addition of 6-azauridine resulted in the accumulation of orotidine, OA, DHO, and carbamyl aspartate (CAA). The use of the observed ratios of [CCA]/[OA] and [DHO]/[OA] and other known constants allowed us to calculate that the increased cellular OA concentration caused primarily an inhibition of DHO dehydrogenase rather than an inhibition of dihydroorotase. A constant ratio of [CAA]/[DHO] was observed which probably indicates that the interconversion of these two intermediates catalyzed by dihydroorotase is near equilibrium in these cells as has been observed in vitro (Christopherson, R.I., Matsuura, T., and Jones, M.E. (1978) Anal. Biochem. 89, 225-234). It is suggested that the probable intracellular accumulation of CAA in patients with oroticaciduria may have significant secondary effects.
D000581:Amidohydrolases / Q000037:antagonists & inhibitors*; D000818:Animals; D001380:Azauridine / Q000494:pharmacology*; D001639:Bicarbonates / Q000378:metabolism; D002286:Carcinoma, Ehrlich Tumor / Q000201:enzymology*; D004080:Dihydroorotase / Q000037:antagonists & inhibitors*; D004081:Dihydroorotate Oxidase / Q000037:antagonists & inhibitors*; D004355:Drug Stability; D007700:Kinetics; D008433:Mathematics; D051379:Mice; D009963:Orotic Acid / Q000494:pharmacology*; D010088:Oxidoreductases / Q000037:antagonists & inhibitors*; D011743:Pyrimidines / Q000096:biosynthesis*
D001639:Bicarbonates; D011743:Pyrimidines; D009963:Orotic Acid; D001380:Azauridine; D010088:Oxidoreductases; D004081:Dihydroorotate Oxidase; D000581:Amidohydrolases; D004080:Dihydroorotase