Cytochrome P450 isozymes catalyzing 4-hydroxylation of parkinsonism-related compound 1,2,3,4-tetrahydroisoquinoline in rat liver microsomes

Tokuji Suzuki, Shoichi Fujita, Shizuo Narimatsu, Yasuhiro Masubuchi, Masaya Tachibana, Shigeru Ohta, Masaaki Hirobe

Research output: Contribution to journalArticle

35 Citations (Scopus)

Abstract

Microsomal 4-hydroxylase of 1,2,3,4-tetrahydroisoquinoline (TIQ), a possible candidate for causing Parkinson disease, was characterized by using rat hepatic microsomes and purified P450 isozymes. Kinetic analysis revealed that Km and Vmax values (mean ± SE) for hepatic microsomal TIQ 4-hydroxylase of male Wistar rats were 319.6 ± 26.8 μM and 12.13 ± 1.43 pmol · min-1 · mg-1 protein, respectively. When TIQ 4-hydroxylase activity was compared in Wistar (an animal model of extensive debrisoquine metabolizers) and Dark Agouti (an animal model of poor debrisoquine metabolizers) rats, significant strain (Wistar > Dark Agouti) and sex (male > female) differences were observed. The microsomal activity toward TIQ 4-hydroxylation was increased by pretreatment of male Wistar rats with P448 inducers (β-naphthoflavone and sudan I), but not with phenobarbital. Pretreatment with propranolol, an inhibitor of P450 isozymes belonging to the P450 IID gene subfamily, decreased TIQ 4-hydroxylase activity. P450 BTL, a P450 isozyme belonging to the IID subfamily, showed TIQ 4-hydroxylase activity of 64.1 pmol · min-1 · nmol P450-1, which was 3.2-fold that of microsomes (20.9 pmol · min-1 · nmol P450-1). Antibody (IgG) against this isozyme suppressed microsomal TIQ 4-hydroxylase activity concentration-dependently. A male-specific P450 ml (P450IIC11) catalyzed this reaction to a much lesser extent (10.0 pmol · min-1 · nmol P450-1), and its antibody did not affect the microsomal activity. These results suggest that TIQ 4-hydroxylation in hepatic microsomes are catalyzed predominantly by a P450 isozyme (or isozymes) belonging to the IID gene subfamily in nontreated rats and its immunochemically related P450 isozyme (or isozymes), and that a P450 isozyme (or isozymes) belonging to the IA subfamily also participates in TIQ 4-hydroxylation in rats pretreated with P448-inducers.

Original languageEnglish
Pages (from-to)771-776
Number of pages6
JournalFASEB Journal
Volume6
Issue number2
Publication statusPublished - Jan 6 1992
Externally publishedYes

Fingerprint

tetrahydroisoquinolines
Hydroxylation
liver microsomes
Parkinsonian Disorders
hydroxylation
Liver Microsomes
cytochrome P-450
Liver
Cytochrome P-450 Enzyme System
Isoenzymes
Rats
isozymes
Mixed Function Oxygenases
rats
Microsomes
Debrisoquin
Agouti
1-phenylazo-2-naphthol
Wistar Rats
Animals

Keywords

  • Dark Agouti rat
  • Debrisoquine
  • Genotypic poor metabolizer
  • Metabolic polymorphism
  • P450 BTL
  • P450 ml
  • Phenotypic poor metabolizer
  • Rat liver microsomes

ASJC Scopus subject areas

  • Agricultural and Biological Sciences (miscellaneous)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Cell Biology

Cite this

Suzuki, T., Fujita, S., Narimatsu, S., Masubuchi, Y., Tachibana, M., Ohta, S., & Hirobe, M. (1992). Cytochrome P450 isozymes catalyzing 4-hydroxylation of parkinsonism-related compound 1,2,3,4-tetrahydroisoquinoline in rat liver microsomes. FASEB Journal, 6(2), 771-776.

Cytochrome P450 isozymes catalyzing 4-hydroxylation of parkinsonism-related compound 1,2,3,4-tetrahydroisoquinoline in rat liver microsomes. / Suzuki, Tokuji; Fujita, Shoichi; Narimatsu, Shizuo; Masubuchi, Yasuhiro; Tachibana, Masaya; Ohta, Shigeru; Hirobe, Masaaki.

In: FASEB Journal, Vol. 6, No. 2, 06.01.1992, p. 771-776.

Research output: Contribution to journalArticle

Suzuki, T, Fujita, S, Narimatsu, S, Masubuchi, Y, Tachibana, M, Ohta, S & Hirobe, M 1992, 'Cytochrome P450 isozymes catalyzing 4-hydroxylation of parkinsonism-related compound 1,2,3,4-tetrahydroisoquinoline in rat liver microsomes', FASEB Journal, vol. 6, no. 2, pp. 771-776.
Suzuki, Tokuji ; Fujita, Shoichi ; Narimatsu, Shizuo ; Masubuchi, Yasuhiro ; Tachibana, Masaya ; Ohta, Shigeru ; Hirobe, Masaaki. / Cytochrome P450 isozymes catalyzing 4-hydroxylation of parkinsonism-related compound 1,2,3,4-tetrahydroisoquinoline in rat liver microsomes. In: FASEB Journal. 1992 ; Vol. 6, No. 2. pp. 771-776.
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AU - Suzuki, Tokuji

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AU - Narimatsu, Shizuo

AU - Masubuchi, Yasuhiro

AU - Tachibana, Masaya

AU - Ohta, Shigeru

AU - Hirobe, Masaaki

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N2 - Microsomal 4-hydroxylase of 1,2,3,4-tetrahydroisoquinoline (TIQ), a possible candidate for causing Parkinson disease, was characterized by using rat hepatic microsomes and purified P450 isozymes. Kinetic analysis revealed that Km and Vmax values (mean ± SE) for hepatic microsomal TIQ 4-hydroxylase of male Wistar rats were 319.6 ± 26.8 μM and 12.13 ± 1.43 pmol · min-1 · mg-1 protein, respectively. When TIQ 4-hydroxylase activity was compared in Wistar (an animal model of extensive debrisoquine metabolizers) and Dark Agouti (an animal model of poor debrisoquine metabolizers) rats, significant strain (Wistar > Dark Agouti) and sex (male > female) differences were observed. The microsomal activity toward TIQ 4-hydroxylation was increased by pretreatment of male Wistar rats with P448 inducers (β-naphthoflavone and sudan I), but not with phenobarbital. Pretreatment with propranolol, an inhibitor of P450 isozymes belonging to the P450 IID gene subfamily, decreased TIQ 4-hydroxylase activity. P450 BTL, a P450 isozyme belonging to the IID subfamily, showed TIQ 4-hydroxylase activity of 64.1 pmol · min-1 · nmol P450-1, which was 3.2-fold that of microsomes (20.9 pmol · min-1 · nmol P450-1). Antibody (IgG) against this isozyme suppressed microsomal TIQ 4-hydroxylase activity concentration-dependently. A male-specific P450 ml (P450IIC11) catalyzed this reaction to a much lesser extent (10.0 pmol · min-1 · nmol P450-1), and its antibody did not affect the microsomal activity. These results suggest that TIQ 4-hydroxylation in hepatic microsomes are catalyzed predominantly by a P450 isozyme (or isozymes) belonging to the IID gene subfamily in nontreated rats and its immunochemically related P450 isozyme (or isozymes), and that a P450 isozyme (or isozymes) belonging to the IA subfamily also participates in TIQ 4-hydroxylation in rats pretreated with P448-inducers.

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