Experimental determination of the phosphorylation state of phenylalanine hydroxylase

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Experimental determination of the phosphorylation state of phenylalanine hydroxylase.

A monoclonal antibody (PH 7), which recognizes the phosphorylated form of phenylalanine hydroxylase from human liver, has been used for the analysis of the enzyme in crude cell extracts from rat. In immunoblot analyses of rat liver cell extracts, the extent of binding of PH 7 closely correlates with the phosphorylation state of phenylalanine hydroxylase, as judged by [32P]Pi incorporation. Thes...

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Phenylalanine hydroxylase (Ec 1.14.16.1) catalyses the initial step of phenylalanine degradation in man and other mammals. Although the liver is the primary location of the enzyme, an appreciable amount is also found in the renal cortex [I]. The regulatory properties of the hepatic enzyme are well known. In the short-term, enzymic activity is controlled by reversible phosphorylation [2]. Glucag...

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Phosphorylation and Mutations of Ser in Human Phenylalanine Hydroxylase

Phosphorylation of phenylalanine hydroxylase (PAH) at Ser by cyclic AMP-dependent protein kinase is a post-translational modification that increases its basal activity and facilitates its activation by the substrate L-Phe. So far there is no structural information on the flexible N-terminal tail (residues 1–18), including the phosphorylation site. To get further insight into the molecular basis...

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Essentially pure phenylalanine hydroxylase from rat liver can be activated between 2.5- and 3.0-fold by treatment with Mg2+, ATP, protein kinase, and cyclic AMP. The activation is seen when the hydroxylase is assayed in the presence of tetrahydrobiopterin, but not in the presence of 2-amino-4-hydroxy-6,7-dimethyltetrahydropteridine. In the presence of [gamma-32P]ATP, activation is accompanied b...

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ژورنال

عنوان ژورنال: Biochemical Journal

سال: 1990

ISSN: 0264-6021,1470-8728

DOI: 10.1042/bj2650563