Tyrosine Hydroxylase Inactivation Following cAMP‐Dependent Phosphorylation Activation

Kent Vrana, Robert Roskoski

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

Tyrosine hydroxylase, the rate‐limiting enzyme in catecholamine biosynthesis, is activated following phosphorylation by the cAMP‐dependent protein kinase (largely by decreasing the Km of the enzyme for its pterin co‐substrate). Following its phosphorylation activation in rat striatal homogenates, we find that tyrosine hydroxylase is inactivated by two distinct processes. Because cAMP is hydrolyzed in crude extracts by a phospho‐diesterase, cAMP‐dependent protein kinase activity declines following a single addition of cAMP. When tyrosine hydroxylase is activated under these transient phosphorylation conditions, inactivation is accompanied by a reversion of the activated kinetic form (low apparent Km for pterin co‐substrate, ≤0.2 mM) to the kinetic form characteristic of the untreated enzyme (high apparent Km, ≥1.0 mM). This inactivation is readily reversed by the subsequent addition of cAMP. When striatal tyrosine hydroxylase is activated under constant phosphorylation conditions (incubated with purified cAMP‐dependent protein kinase catalytic subunit), however, it is also inactivated. This second inactivation process is irreversible and is characterized kinetically by a decreasing apparent Vmax with no change in the low apparent Km for pterin co‐substrate (0.2 mM). The latter inactivation process is greatly attenuated by gel filtration which resolves a low‐molecular‐weight inactivating factor(s) from the tyrosine hydroxylase. These results are consistent with a regulatory mechanism for tyrosine hydroxylase involving two processes: in the first case, reversible phosphorylaton and dephos‐phorylation and, in the second case, an irreversible loss of activity of the phosphorylated form of tyrosine hydroxylase.

Original languageEnglish (US)
Pages (from-to)1692-1700
Number of pages9
JournalJournal of Neurochemistry
Volume40
Issue number6
DOIs
StatePublished - Jan 1 1983

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Phosphorylation
Tyrosine 3-Monooxygenase
Chemical activation
Pterins
Protein Kinases
Corpus Striatum
Enzymes
Kinetics
Biosynthesis
Phosphoric Diester Hydrolases
Complex Mixtures
Catecholamines
Gel Chromatography
Rats
Catalytic Domain
Gels

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Cellular and Molecular Neuroscience

Cite this

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title = "Tyrosine Hydroxylase Inactivation Following cAMP‐Dependent Phosphorylation Activation",
abstract = "Tyrosine hydroxylase, the rate‐limiting enzyme in catecholamine biosynthesis, is activated following phosphorylation by the cAMP‐dependent protein kinase (largely by decreasing the Km of the enzyme for its pterin co‐substrate). Following its phosphorylation activation in rat striatal homogenates, we find that tyrosine hydroxylase is inactivated by two distinct processes. Because cAMP is hydrolyzed in crude extracts by a phospho‐diesterase, cAMP‐dependent protein kinase activity declines following a single addition of cAMP. When tyrosine hydroxylase is activated under these transient phosphorylation conditions, inactivation is accompanied by a reversion of the activated kinetic form (low apparent Km for pterin co‐substrate, ≤0.2 mM) to the kinetic form characteristic of the untreated enzyme (high apparent Km, ≥1.0 mM). This inactivation is readily reversed by the subsequent addition of cAMP. When striatal tyrosine hydroxylase is activated under constant phosphorylation conditions (incubated with purified cAMP‐dependent protein kinase catalytic subunit), however, it is also inactivated. This second inactivation process is irreversible and is characterized kinetically by a decreasing apparent Vmax with no change in the low apparent Km for pterin co‐substrate (0.2 mM). The latter inactivation process is greatly attenuated by gel filtration which resolves a low‐molecular‐weight inactivating factor(s) from the tyrosine hydroxylase. These results are consistent with a regulatory mechanism for tyrosine hydroxylase involving two processes: in the first case, reversible phosphorylaton and dephos‐phorylation and, in the second case, an irreversible loss of activity of the phosphorylated form of tyrosine hydroxylase.",
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Tyrosine Hydroxylase Inactivation Following cAMP‐Dependent Phosphorylation Activation. / Vrana, Kent; Roskoski, Robert.

In: Journal of Neurochemistry, Vol. 40, No. 6, 01.01.1983, p. 1692-1700.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Tyrosine Hydroxylase Inactivation Following cAMP‐Dependent Phosphorylation Activation

AU - Vrana, Kent

AU - Roskoski, Robert

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Y1 - 1983/1/1

N2 - Tyrosine hydroxylase, the rate‐limiting enzyme in catecholamine biosynthesis, is activated following phosphorylation by the cAMP‐dependent protein kinase (largely by decreasing the Km of the enzyme for its pterin co‐substrate). Following its phosphorylation activation in rat striatal homogenates, we find that tyrosine hydroxylase is inactivated by two distinct processes. Because cAMP is hydrolyzed in crude extracts by a phospho‐diesterase, cAMP‐dependent protein kinase activity declines following a single addition of cAMP. When tyrosine hydroxylase is activated under these transient phosphorylation conditions, inactivation is accompanied by a reversion of the activated kinetic form (low apparent Km for pterin co‐substrate, ≤0.2 mM) to the kinetic form characteristic of the untreated enzyme (high apparent Km, ≥1.0 mM). This inactivation is readily reversed by the subsequent addition of cAMP. When striatal tyrosine hydroxylase is activated under constant phosphorylation conditions (incubated with purified cAMP‐dependent protein kinase catalytic subunit), however, it is also inactivated. This second inactivation process is irreversible and is characterized kinetically by a decreasing apparent Vmax with no change in the low apparent Km for pterin co‐substrate (0.2 mM). The latter inactivation process is greatly attenuated by gel filtration which resolves a low‐molecular‐weight inactivating factor(s) from the tyrosine hydroxylase. These results are consistent with a regulatory mechanism for tyrosine hydroxylase involving two processes: in the first case, reversible phosphorylaton and dephos‐phorylation and, in the second case, an irreversible loss of activity of the phosphorylated form of tyrosine hydroxylase.

AB - Tyrosine hydroxylase, the rate‐limiting enzyme in catecholamine biosynthesis, is activated following phosphorylation by the cAMP‐dependent protein kinase (largely by decreasing the Km of the enzyme for its pterin co‐substrate). Following its phosphorylation activation in rat striatal homogenates, we find that tyrosine hydroxylase is inactivated by two distinct processes. Because cAMP is hydrolyzed in crude extracts by a phospho‐diesterase, cAMP‐dependent protein kinase activity declines following a single addition of cAMP. When tyrosine hydroxylase is activated under these transient phosphorylation conditions, inactivation is accompanied by a reversion of the activated kinetic form (low apparent Km for pterin co‐substrate, ≤0.2 mM) to the kinetic form characteristic of the untreated enzyme (high apparent Km, ≥1.0 mM). This inactivation is readily reversed by the subsequent addition of cAMP. When striatal tyrosine hydroxylase is activated under constant phosphorylation conditions (incubated with purified cAMP‐dependent protein kinase catalytic subunit), however, it is also inactivated. This second inactivation process is irreversible and is characterized kinetically by a decreasing apparent Vmax with no change in the low apparent Km for pterin co‐substrate (0.2 mM). The latter inactivation process is greatly attenuated by gel filtration which resolves a low‐molecular‐weight inactivating factor(s) from the tyrosine hydroxylase. These results are consistent with a regulatory mechanism for tyrosine hydroxylase involving two processes: in the first case, reversible phosphorylaton and dephos‐phorylation and, in the second case, an irreversible loss of activity of the phosphorylated form of tyrosine hydroxylase.

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