Cloning of a cDNA encoding a novel marmoset CYP2C enzyme, expression in yeast cells and characterization of its enzymatic functions

Shizuo Narimatsu, Fumihiro Torigoe, Yumi Tsuneto, Keita Saito, Nobumitsu Hanioka, Kazufumi Masuda, Takashi Katsu, Shigeo Yamamoto, Shigeru Yamano, Takahiko Baba, Atsuro Miyata

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

We cloned a cDNA encoding a novel CYP2C enzyme, called P450 M-2C, from a marmoset liver. The deduced amino acid sequence showed high identities to those of human CYP2C8 (87%), CYP2C9 (78%) and CYP2C19 (77%). The P450 M-2C enzyme expressed in yeast cells catalyzed p-methylhydroxylation of only tolbutamide among four substrates tested, paclitaxel as a CYP2C8 substrate, diclofenac and tolbutamide as CYP2C9 substrates and S-mephenytoin as a CYP2C19 substrate. p-Methylhydroxylation of tolbutamide by marmoset liver microsomes showed monophasic kinetics, and the apparent Km value (1.2 mM) for the substrate was similar to that of the recombinant P450 M-2C (1.8 mM). Although all of the recombinant human CYP2C8, CYP2C9 and CYP2C19 expressed in yeast cells catalyzed tolbutamide p-methylhydroxylation, the kinetic profile of CYP2C8 was most similar to that of P450 M-2C. Tolbutamide oxidation by the marmoset liver microsomes and the recombinant P450 M-2C was inhibited most effectively by quercetin, a CYP2C8 inhibitor, followed by omeprazole, a CYP2C19 inhibitor, whereas sulfaphenazole, a CYP2C9 inhibitor, was less potent under the conditions used. These results indicate that P450 M-2C is the major tolbutamide p-methylhydroxylase in the marmoset liver.

Original languageEnglish
Pages (from-to)1738-1748
Number of pages11
JournalBiochemical Pharmacology
Volume72
Issue number12
DOIs
Publication statusPublished - Dec 15 2006
Externally publishedYes

Keywords

  • CYP2C8
  • Marmoset
  • Quercetin
  • Tolbutamide
  • Yeast cell

ASJC Scopus subject areas

  • Biochemistry
  • Pharmacology

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    Narimatsu, S., Torigoe, F., Tsuneto, Y., Saito, K., Hanioka, N., Masuda, K., Katsu, T., Yamamoto, S., Yamano, S., Baba, T., & Miyata, A. (2006). Cloning of a cDNA encoding a novel marmoset CYP2C enzyme, expression in yeast cells and characterization of its enzymatic functions. Biochemical Pharmacology, 72(12), 1738-1748. https://doi.org/10.1016/j.bcp.2006.08.025