title: Phosphorylation and activation of brain aromatic L-amino acid decarboxylase by cyclic AMP-dependent protein kinase.
abstract: Aromatic L-amino acid decarboxylase (AAAD), an enzyme required for the synthesis of catecholamines, indoleamines, and trace amines, is rapidly activated by cyclic AMP-dependent pathways in striatum and midbrain in vivo, suggesting enzyme phosphorylation. We now report that the catalytic subunit of cyclic AMP-dependent protein kinase (PKA) directly phosphorylated AAAD immunoprecipitated from homogenates prepared from the mouse striatum and midbrain in vitro. Under the same phosphorylation conditions, the catalytic subunit of PKA also phosphorylated a recombinant AAAD protein expressed in Escherichia coli transfected with an AAAD cDNA isolated from the bovine adrenal gland. The PKA-induced AAAD phosphorylation of immunoprecipitates from striatum and midbrain was time and concentration dependent and blocked by a specific PKA peptide inhibitor. Incubation of the catalytic subunit of PKA with striatal homogenates increased enzyme activity by approximately 20% in a time- and concentration-dependent manner. Moreover, incubation of the catalytic subunit of PKA with recombinant AAAD increased activity by approximately 70%. A direct phosphorylation of AAAD protein by PKA might underlie the cyclic AMP-induced rapid and transient activation of AAAD in vivo.
authors.affiliation: Department of Psychiatry, Ohio State University College of Medicine and Public Health, Columbus, Ohio, USA.
authors.forename: A M
authors.initials: AM
authors.lastname: Duchemin
authors.forename: M D
authors.initials: MD
authors.lastname: Berry
authors.forename: N H
authors.initials: NH
authors.lastname: Neff
authors.forename: M
authors.initials: M
authors.lastname: Hadjiconstantinou
mesh_terms: D000311:Adrenal Glands / Q000201:enzymology; D000818:Animals; D001142:Aromatic-L-Amino-Acid Decarboxylases / Q000378:metabolism*; D001921:Brain / Q000201:enzymology*; D002417:Cattle; D003001:Cloning, Molecular; D003342:Corpus Striatum / Q000201:enzymology; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism*; D004789:Enzyme Activation; D004926:Escherichia coli; D007700:Kinetics; D008297:Male; D008636:Mesencephalon / Q000201:enzymology; D051379:Mice; D010766:Phosphorylation; D011994:Recombinant Proteins / Q000378:metabolism
chemical_list: D011994:Recombinant Proteins; D017868:Cyclic AMP-Dependent Protein Kinases; D001142:Aromatic-L-Amino-Acid Decarboxylases