Regulation of phenylalanine hydroxylase activity by phenylalanine in vivo, in vitro, and in perfused rat liver.
We show that phenylalanine is able to control the extent of activation and, as a result, the catalytic activity of rat liver phenylalanine hydroxylase in vivo, in perfused liver, and in vitro. Both phosphorylated and unphosphorylated enzyme activities are controlled by phenylalanine activation and, overall, this mechanism appears to be a major means of regulating the enzyme's activity in rat liver. At normal phenylalanine levels in vivo, phenylalanine hydroxylase is at most 1-4% activated, and phosphorylated enzyme (glucagon-induced) appears at most 5-7% activated under similar conditions. In both cases, a phenylalanine load increased the percentage of activated enzyme found in vivo to about 40% of maximal. In perfused rat livers, a plasma phenylalanine concentration of only 4 times normal induced a 4-fold increase in the amount of activated enzyme present and a corresponding functional increase in the rate of phenylalanine hydroxylation by the tissue. Under the latter conditions, more than 25% of the amino acid could be hydroxylated in a single pass through the organ. Purified phosphorylated phenylalanine hydroxylase must be activated to be catalytically active. The activation with phenylalanine, at equilibrium, is a cooperative process, and the phosphorylated enzyme is activated more rapidly at pH 6.8 and 8.0 and at lower phenylalanine concentration than the unphosphorylated species. Overall, phosphorylation appears to allow phenylalanine hydroxylase to be more easily activated at relatively low phenylalanine concentrations.
D000818:Animals; D004789:Enzyme Activation; D005934:Glucagon / Q000494:pharmacology; D007700:Kinetics; D008099:Liver / Q000187:drug effects / Q000201:enzymology*; D008297:Male; D010477:Perfusion; D010649:Phenylalanine / Q000494:pharmacology*; D010651:Phenylalanine Hydroxylase / Q000378:metabolism*; D051381:Rats; D011917:Rats, Inbred Lew
D010649:Phenylalanine; D005934:Glucagon; D010651:Phenylalanine Hydroxylase