Mutation of regulatory serines of rat tyrosine hydroxylase to glutamate: Effects on enzyme stability and activity

Montserrat Royo, Paul F. Fitzpatrick, S. Colette Daubner

Research output: Contribution to journalArticle

25 Scopus citations

Abstract

Tyrosine hydroxylase is phosphorylated at four serine residues in its amino-terminus by multiple kinases. Phosphorylation of serine 40 by cAMP-dependent protein kinase results in alleviation of dopamine inhibition [J. Biol. Chem. 267 (1992) 12639]. The other serines are at positions 8, 19, and 31. The effect of phosphorylation at these serines has been investigated using mutated forms of tyrosine hydroxylase containing glutamates at the positions of the serines. The S8E, S19E, and S31E tyrosine hydroxylase variants have similar steady-state kinetic parameters and similar binding affinity for catecholamines to wild-type enzyme. The S8E, S19E, S31E, and S40E variants differ in stability at elevated temperatures. The S40E variant is the least stable, while the others are all more stable than wild-type enzyme. The increased stability of S8E, S19E, and S31E tyrosine hydroxylases may be one of the physiological effects of phosphorylation. It may also have implications for the interpretation of activities of heterogeneous mixtures of tyrosine hydroxylase which have been phosphorylated.

Original languageEnglish (US)
Pages (from-to)266-274
Number of pages9
JournalArchives of Biochemistry and Biophysics
Volume434
Issue number2
DOIs
Publication statusPublished - Feb 15 2005

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Keywords

  • Dopamine
  • Enzyme phosphorylation
  • Enzyme regulation
  • Enzyme stability
  • Site-directed mutagenesis
  • Tyrosine hydroxylase

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology

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