Characterization of Recombinant Human Aromatic l‐Amino Acid Decarboxylase Expressed in COS Cells

Chiho Ichinose, Hiroshi Ichinose, Toshiharu Nagatsu

Research output: Contribution to journalArticle

34 Citations (Scopus)

Abstract

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 coprecip‐itation 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.

Original languageEnglish
Pages (from-to)1075-1078
Number of pages4
JournalJournal of Neurochemistry
Volume55
Issue number3
DOIs
Publication statusPublished - 01-01-1990

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Carboxy-Lyases
COS Cells
Carboxylic acids
Complementary DNA
Pyridoxal Phosphate
Acids
Enzyme activity
Substrates
Enzymes
Coprecipitation
Decarboxylation
Plasmids
Direction compound
calcium phosphate

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Cellular and Molecular Neuroscience

Cite this

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abstract = "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 coprecip‐itation 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.",
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Characterization of Recombinant Human Aromatic l‐Amino Acid Decarboxylase Expressed in COS Cells. / Ichinose, Chiho; Ichinose, Hiroshi; Nagatsu, Toshiharu.

In: Journal of Neurochemistry, Vol. 55, No. 3, 01.01.1990, p. 1075-1078.

Research output: Contribution to journalArticle

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AU - Ichinose, Hiroshi

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AB - 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 coprecip‐itation 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.

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