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Rafii M, McKenzie JM, Roberts SA, Steiner G, Ball RO, Pencharz PB: In vivo regulation of phenylalanine hydroxylation to tyrosine, studied using enrichment in apoB-100. Am J Physiol Endocrinol Metab. 2008 Feb;294(2):E475-9. Epub 2007 Nov 27.
Phenylalanine hydroxylation is necessary for the conversion of phenylalanine to tyrosine and disposal of excess phenylalanine. Studies of in vivo regulation of phenylalanine hydroxylation suffer from the lack of a method to determine intrahepatocyte enrichment of phenylalanine and tyrosine. apoB-100, a hepatic export protein, is synthesized from intrahepatocyte amino acids. We designed an in vivo multi-isotope study, [(15) N] phenylalanine and [2H2] tyrosine to determine rates of phenylalanine hydroxylation from plasma enrichments in free amino acids and apoB-100. For independent verification of apoB-100 as a reflection of enrichment in the intrahepatocyte pool, [1-(13) C] lysine was used as an indicator amino acid (IAA) to measure in vivo changes in protein synthesis in response to tyrosine supplementation. Adult men (n = 6) were fed an amino acid-based diet with low phenylalanine (9 mg.kg (-1).day (-1), 4.54 mumol.kg (-1).,h (-1)) and seven graded intakes of tyrosine from 2.5 (deficient) to 12.5 (excess) mg.kg (-1).day (-1). Gas chromatography-quadrupole mass spectrometry did not detect any tracer in apoB-100 tyrosine. A new and more sensitive method to measure label enrichment in proteins using isotope ratio mass spectrometry demonstrated that phenylalanine hydroxylation measured in apoB-100 decreased linearly in response to increasing tyrosine intake and reached a break point at 6.8 mg.kg (-1).day (-1). IAA oxidation decreased with increased tyrosine intake and reached a break point at 6.0 mg.kg (-1).day (-1). We conclude: apoB-100 is an accurate and useful measure of changes in phenylalanine hydroxylation; the synthesis of tyrosine via phenylalanine hydroxylation is regulated to meet the needs for protein synthesis; and that plasma phenylalanine does not reflect changes in protein synthesis. |
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