Studies on the interaction of a thiol-dependent hydrogen peroxide scavenging enzyme and phenylalanine hydroxylase.
Rat liver phenylalanine hydroxylase is irreversibly inactivated by a H2O2-dependent process. Since H2O2 can be produced by autooxidation of the tetrahydropterin cofactor required for the hydroxylation reaction, in vitro assays are usually carried out in the presence of added catalase. On the basis of a dithiothreitol-dependent protecting assay of phenylalanine hydroxylase, carried out in the absence of catalase, we have isolated an enzyme fraction from neonatal rat livers which has similar properties to the known enzyme, glutathione peroxidase. The developmental time course for phenylalanine hydroxylase in rats has been reported to follow two different patterns. Using the dithiothreitol assay, McGee et al. (1972, Biochem. J. 127, 669-674) have found that newborn rats have low phenylalanine hydroxylase activity which increases to adult levels over several months. On the other hand, using catalase-supplemented assays, others have found that newborn rats have nearly adult levels of phenylalanine hydroxylase activity. The protective effect of glutathione peroxidase on phenylalanine hydroxylase suggests that the developmental time course found by McGee et al. may represent the slow developmental time course previously found for glutathione peroxidase. In addition, feeding rats a selenium-deficient diet, which reduces the hepatic activity of the selenium-containing glutathione peroxidase, results in a concomitant irreversible loss of phenylalanine hydroxylase activity, suggesting that glutathione peroxidase may play a vital role in protecting phenylalanine hydroxylase in vivo from peroxide inactivation.
Milstien
S
S
Laboratory of Neurochemistry, National Institute of Mental Health, Bethesda, Maryland 20892.
Dorche
C
C
Kaufman
S
S
D000818:Animals; D004347:Drug Interactions; D005979:Glutathione Peroxidase / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism*; D006861:Hydrogen Peroxide / Q000378:metabolism* / Q000494:pharmacology; D008099:Liver / Q000201:enzymology*; D010651:Phenylalanine Hydroxylase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains; D012643:Selenium / Q000172:deficiency; D013379:Substrate Specificity
D006861:Hydrogen Peroxide; D005979:Glutathione Peroxidase; D010651:Phenylalanine Hydroxylase; D012643:Selenium