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By Seymour Kaufman

The severely acclaimed laboratory ordinary, Methods in Enzymology, is without doubt one of the such a lot hugely revered guides within the box of biochemistry. for the reason that 1955, each one quantity has been eagerly awaited, often consulted, and praised by means of researchers and reviewers alike. The sequence comprises a lot fabric nonetheless proper this present day - actually a vital booklet for researchers in all fields of existence sciences

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The assay sensitivity can be greatly increased by substituting radiolabeled phenylalanine and isolating the labeled tyrosine formed. H,n 4. 4× its Kin. If desired, 120/zM 6-methyltetrahydropterin can be substituted. 3 x increase in the apparent specific activity of the enzyme. 5. This assay has been used with no problem on crude extracts of rabbit, rat, monkey, and goose liver. However, since tyrosine formation in the assay is calculated from an absorbance change, when a new source of enzyme is examined, this absorbance assay should always be confirmed by a method (such as the radiochemical procedures cited above) that directly measures phenylalanine conversion to tyrosine.

2] PHENYLALANINE 4-MONOt~XYGENASE (RAT LIVER) 17 and with the pterin used to assay its activity. 0. 0. 52 55 M. Parniak and S. Kaufman,Biochemistry 24, 3379 (abstr. No. 133) (1985). [2] P u r i f i c a t i o n a n d A s s a y o f R a t L i v e r P h e n y l a l a n i n e 4-Monooxygenase By Ross SH1MAN Mammalian phenylalanine 4-monooxygenase (hydroxylase) catalyzes the hydroxylation of phenylalanine to tyrosine. The enzyme is soluble and cytoplasmic and is found mainly in liver. The hydroxylation reaction is irreversible and requires Oz and an unconjugated tetrahydropterin as cosubstrates [Eq.

Several catechols that are intermediates in the conversion of tyrosine to epinephrine--dopa, dopamine, and norepinephrine--inhibit the enzyme with Ki values of about l0 -5 M. 49 Metal chelating agents, such as 2,2'-dipyridine, o-phenanthroline, 8-hydroxyquinoline, and diethyldithiocarbamate inhibit, whereas EDTA (up to 20 mM) does not. nes, such as the p-chloro derivative, 5° are moderate inhibitors of phenylalanine hydroxylation. Other Properties. , it is in the 100,000 g supernatant fraction obtained from homogenates prepared with isotonic sucrose or salt solutions.

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