title: Distinct forms of the protein kinase-dependent activator of tyrosine and tryptophan hydroxylases.
abstract: Tyrosine and tryptophan hydroxylases are the key enzymes in the regulation of catecholamine and serotonin levels in neurons and other endocrine cells. Among the mechanisms proposed for the modulation of activity, phosphorylation of the enzyme is believed to be of functional significance with respect to the stimulus-response coupling, but the precise mechanism is unknown. Here, we show the existence of multiple, distinct forms of the 14-3-3 activator protein, a neuronal protein essential for activation of tyrosine and tryptophan hydroxylases by Ca2+/calmodulin-dependent protein kinase type II. Bovine brain 14-3-3 protein was resolved by reversed-phase chromatography into seven polypeptides (alpha to eta), all of which were active towards tryptophan hydroxylase when the renatured preparations were assayed in the presence of Ca2+, calmodulin and the protein kinase. Determination of the amino acid sequences of the beta and gamma chains and comparison of the sequences with the previously determined sequence of the eta chain revealed that these molecules are highly homologous, and share a common structural feature in containing an extremely acidic C-terminal region predicted as a domain for interaction with the phosphorylated hydroxylases. Northern blot analysis indicated that the beta, gamma and eta chain are expressed abundantly in the brain; however, these polypeptides appear to be expressed with different tissue specificities because gamma mRNA is found only in the brain, while lower levels of beta and eta mRNAs are detected in several other tissues. These findings suggest the involvement of a diverse family of the activator protein in the stimulus-coupled, Ca2(+)-dependent regulation of monoamine biosynthesis.
authors.affiliation: Department of Chemistry, Faculty of Science, Tokyo Metropolitan University, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Isobe
authors.forename: T
authors.initials: T
authors.lastname: Ichimura
authors.forename: T
authors.initials: T
authors.lastname: Sunaya
authors.forename: T
authors.initials: T
authors.lastname: Okuyama
authors.forename: N
authors.initials: N
authors.lastname: Takahashi
authors.forename: R
authors.initials: R
authors.lastname: Kuwano
authors.forename: Y
authors.initials: Y
authors.lastname: Takahashi
mesh_terms: D048948:14-3-3 Proteins; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015306:Biogenic Monoamines / Q000096:biosynthesis; D015152:Blotting, Northern; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D004789:Enzyme Activation; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011494:Protein Kinases / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D012689:Sequence Homology, Nucleic Acid; D014365:Tryptophan Hydroxylase / Q000378:metabolism*; D014446:Tyrosine 3-Monooxygenase / Q000378:metabolism*
chemical_list: D048948:14-3-3 Proteins; D015306:Biogenic Monoamines; D009419:Nerve Tissue Proteins; D012333:RNA, Messenger; D014446:Tyrosine 3-Monooxygenase; D014365:Tryptophan Hydroxylase; D011494:Protein Kinases