Tyrosine hydroxylase assay for detection of low levels of enzyme activity in peripheral tissues

Douglas Hooper, Minoru Kawamura, Beth Hoffman, Irwin J. Kopin, Bela Hunyady, Eva Mezey, Graeme Eisenhofer

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

A nonisotopic assay for tyrosine hydroxylase, with optimized signal-to-noise ratios, enables determination of low levels of enzyme activity in peripheral tissues. DOPA produced by the enzyme is measured using HPLC with electrochemical detection, increased signal-to-noise ratios are obtained by including in the reaction mixture glycerol for reduction of blank values and dihydropteridine reductase and NADPH for regeneration of the tetrahydropteridine cofactor. With this method, tyrosine hydroxylase activity can be detected in as few as 200 PC12 cells and in peripheral tissues at levels as low as 4.5 fmol/min/mg wet weight, The assay permits activity to be assessed in a variety of peripheral tissues.

Original languageEnglish
Pages (from-to)317-324
Number of pages8
JournalJournal of Chromatography B: Biomedical Applications
Volume694
Issue number2
DOIs
Publication statusPublished - Jul 4 1997

Keywords

  • DOPA
  • Enzymes
  • Tyrosine hydroxylase

ASJC Scopus subject areas

  • Chemistry(all)

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