Products of the tyrosine-dependent oxidation of tetrahydrobiopterin by rat liver phenylalanine hydroxylase.
Phenylalanine hydroxylase, a tetrahydrobiopterin (BH4)-dependent oxygenase, catalyzes the conversion of phenylalanine to tyrosine. During this physiological reaction, the oxidation of BH4 is tightly coupled to the hydroxylation of the amino acid substrate with a stoichiometry of 1 mol of BH4 oxidized for every mole of tyrosine formed. In the presence of an activator and certain analogues of either the amino acid or the pterin coenzyme, the enzyme also catalyzes a partially uncoupled reaction in which the amount of tetrahydropterin oxidized exceeds the amount of amino acid hydroxylated. With tyrosine as the amino acid analogue, the reaction is completely uncoupled, i.e., the enzyme functions as a tetrahydropterin oxidase with no net hydroxylation of the amino acid. We have investigated whether the normal pterin intermediate, a 4a-hydroxytetrahydropterin, is formed during the completely uncoupled reaction and whether oxygen is reduced exclusively to the level of water, as in the tightly coupled reaction, or whether some reduction to hydrogen peroxide occurs. We present direct evidence that both the normal hydroxytetrahydropterin intermediate and hydrogen peroxide are formed. The amount of hydrogen peroxide formed, however, is not stoichiometric with the amount of oxygen reduced, an indication that there are at least two pathways for this reduction, one leading to the formation of water and one to hydrogen peroxide. We also postulate that hydrogen peroxide and the hydroxytetrahydropterin are both derived from alternate routes of breakdown of a common precursor, the corresponding 4a-hydroperoxytetrahydropterin. By contrast, in the normal tightly coupled reaction, we suggest that this peroxy compound participates in the hydroxylation of the amino acid substrate.
Davis
M D
MD
Laboratory of Neurochemistry, National Institute of Mental Health, Bethesda, Maryland 20892.
Kaufman
S
S
D000818:Animals; D001708:Biopterins / Q000031:analogs & derivatives* / Q000378:metabolism; D006861:Hydrogen Peroxide / Q000378:metabolism; D008099:Liver / Q000201:enzymology; D010084:Oxidation-Reduction; D010651:Phenylalanine Hydroxylase / Q000378:metabolism*; D051381:Rats; D014443:Tyrosine / Q000378:metabolism
D001708:Biopterins; D014443:Tyrosine; D006861:Hydrogen Peroxide; D010651:Phenylalanine Hydroxylase; C003402:sapropterin