Molecular cloning and functional analysis of cytochrome P450 1A2 from Japanese monkey liver

Comparison with marmoset cytochrome P450 1A2

Shizuo Narimatsu, Maiko Oda, Hiroyuki Hichiya, Takashi Isobe, Kazuo Asaoka, Nobumitsu Hanioka, Shigeru Yamano, Sumio Shinoda, Shigeo Yamamoto

Research output: Contribution to journalArticle

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Abstract

A cDNA encoding a novel cytochrome P450 1A2 (CYP1A2) was cloned from the liver of an adult female Japanese monkey. The CYP1A2 protein was expressed in yeast cells and its enzymatic properties were compared with those of marmoset CYP1A2 using ethoxyresorufin (ER) and phenacetin (PN) as substrates. The nucleotide sequence of Japanese monkey CYP1A2 revealed 94.7, 99.5 and 93.5% identities to those of human, cynomolgus monkey and marmoset monkey CYP1A2, respectively. Multiple amino acid sequence alignment of Japanese monkey CYP1A2 with CYP1A2 of humans, cynomolgus monkeys and marmosets showed that Japanese monkey CYP1A2 had 92.4, 99.0 and 91.9% identities to the human, cynomolgus monkey and marmoset enzymes, respectively. Kinetic studies demonstrated that the enzymatic properties as ER and PN O-deethylases were considerably different between the Japanese monkey and the marmoset CYP1A2. Furthermore, both of these reactions in liver microsomal fractions from the Japanese monkey and marmoset showed biphasic kinetics. On the basis of the kinetic parameters, it is suggested that Japanese monkey CYP1A2 is a high-Km enzyme in both ER and PN O-deethylations, whereas marmoset CYP1A2 is a high-Km and low-Km enzyme in ER and PN O-deethylations, respectively. α-Naphthoflavone, an inhibitor of human CYP1A1 and CYP1A2, did not completely inhibit the liver microsomal oxidations of ER and PN even at the highest concentration (50 μM), supporting the notion that CYP1A2 enzymes are not the sole ER or PN O-deethylase in Japanese monkey and marmoset liver microsomes. Inhibitory effects of furafylline, an inhibitor of human CYP1A2, on ER O-deethylation by recombinant CYP1A2 enzymes were much lower than those of α-naphthoflavone, but marmoset CYP1A2 was more sensitive to furafylline than Japanese monkey CYP1A2. These results indicate that the properties of Japanese monkey CYP1A2 are considerably different from those of marmoset CYP1A2.

Original languageEnglish
Pages (from-to)1-12
Number of pages12
JournalChemico-Biological Interactions
Volume152
Issue number1
DOIs
Publication statusPublished - Feb 28 2005

Fingerprint

Callithrix
Cytochrome P-450 CYP1A2
Functional analysis
Cloning
Macaca
Molecular Cloning
Liver
Phenacetin
Macaca fascicularis
Cytochrome P-450 CYP1A1
Cytochrome P-450 Enzyme System
Enzymes

Keywords

  • Cytochrome P450 1A2
  • Ethoxyresorufin
  • Japanese monkey
  • Marmoset
  • Phenacetin

ASJC Scopus subject areas

  • Toxicology

Cite this

Molecular cloning and functional analysis of cytochrome P450 1A2 from Japanese monkey liver : Comparison with marmoset cytochrome P450 1A2. / Narimatsu, Shizuo; Oda, Maiko; Hichiya, Hiroyuki; Isobe, Takashi; Asaoka, Kazuo; Hanioka, Nobumitsu; Yamano, Shigeru; Shinoda, Sumio; Yamamoto, Shigeo.

In: Chemico-Biological Interactions, Vol. 152, No. 1, 28.02.2005, p. 1-12.

Research output: Contribution to journalArticle

Narimatsu, S, Oda, M, Hichiya, H, Isobe, T, Asaoka, K, Hanioka, N, Yamano, S, Shinoda, S & Yamamoto, S 2005, 'Molecular cloning and functional analysis of cytochrome P450 1A2 from Japanese monkey liver: Comparison with marmoset cytochrome P450 1A2', Chemico-Biological Interactions, vol. 152, no. 1, pp. 1-12. https://doi.org/10.1016/j.cbi.2005.01.006
Narimatsu, Shizuo ; Oda, Maiko ; Hichiya, Hiroyuki ; Isobe, Takashi ; Asaoka, Kazuo ; Hanioka, Nobumitsu ; Yamano, Shigeru ; Shinoda, Sumio ; Yamamoto, Shigeo. / Molecular cloning and functional analysis of cytochrome P450 1A2 from Japanese monkey liver : Comparison with marmoset cytochrome P450 1A2. In: Chemico-Biological Interactions. 2005 ; Vol. 152, No. 1. pp. 1-12.
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AU - Isobe, Takashi

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