Characterization of recombinant human aromatic L-amino acid decarboxylase expressed in COS cells

J Neurochem. 1990 Sep;55(3):1075-8. doi: 10.1111/j.1471-4159.1990.tb04601.x.

Abstract

The expression vector containing the full-length cDNA of human aromatic L-amino acid decarboxylase (EC 4.1.1.28) was transfected in COS cells by a modified calcium phosphate coprecipitation method. The cells transfected with plasmids that had a true direction of the cDNA gave a major immunoreactive band at 50 kDa. This expressed enzyme catalyzed the decarboxylation of L-3,4-dihydroxyphenylalanine (L-DOPA), L-5-hydroxytryptophan (L-5-HTP) and L-threo-3,4-dihydroxyphenylserine. The optimal pH of the enzyme activity with L-DOPA as a substrate was 6.5, whereas the enzyme had a broad pH optimum when L-5-HTP was used as a substrate. Addition of pyridoxal phosphate to the incubation mixture greatly enhanced the activity for both L-DOPA and L-5-HTP.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 5-Hydroxytryptophan / metabolism
  • Aromatic-L-Amino-Acid Decarboxylases / genetics
  • Aromatic-L-Amino-Acid Decarboxylases / metabolism*
  • Cell Line
  • DNA / genetics
  • Dihydroxyphenylalanine / metabolism
  • Droxidopa / metabolism
  • Humans
  • Hydrogen-Ion Concentration
  • Plasmids
  • Pyridoxal Phosphate / pharmacology
  • Recombinant Proteins / metabolism*
  • Substrate Specificity
  • Transfection

Substances

  • Recombinant Proteins
  • Pyridoxal Phosphate
  • Dihydroxyphenylalanine
  • DNA
  • 5-Hydroxytryptophan
  • Aromatic-L-Amino-Acid Decarboxylases
  • Droxidopa