Regular Article
Molecular Cloning, Expression, and Characterization of CYP2D17 from Cynomolgus Monkey Liver

https://doi.org/10.1006/abbi.1999.1506Get rights and content

Abstract

The cynomolgus monkey is a species used in drug-safety evaluation and biotransformation studies by the pharmaceutical industry. Relatively little is known, however, about the catalytic activities and specificities of cytochromes P450 (CYP) in this species. As a first step in characterizing monkey CYPs, a cDNA was cloned by reverse-transcriptase PCR from cynomolgus monkey liver mRNA using oligonucleotide primers based on the human CYP2D6 sequence. The full-length cDNA (called CYP2D17) encoded a 497-amino-acid protein that is 93% identical to human CYP2D6 and 90% identical to marmoset CYP2D19. The CYP2D17 cDNA was cloned into a baculovirus expression vector, and microsomes prepared from CYP2D17-infected insect cells were used to determine the catalytic properties of the recombinant enzyme. The recombinant CYP2D17 results were compared to data generated with monkey liver microsomes, human liver microsomes, and recombinant CYP2D6 and demonstrated catalytic similarity using probe substrates and inhibitors. Recombinant CYP2D17 catalyzed the oxidation of bufuralol to 1′-hydroxybufuralol and dextromethorphan to dextrorphan, reactions shown to be mediated by CYP2D6 in humans; the apparent Km values for bufuralol and dextromethorphan were 1 and 0.8 μM, respectively. Moreover, both of these reactions were more strongly inhibited by quinidine than by quinine. A more complete understanding of the substrate specificities and activities of monkey CYPs will be advantageous in delineating species differences in metabolite profiles and metabolic activation of new chemical entities in the pharmaceutical industry.

References (35)

  • M. Kastner et al.

    Biochem. Biophys. Acta

    (1994)
  • K. Ohta et al.

    Biochem. Biophys. Acta

    (1989)
  • T.G. Schulz et al.

    Biochim. Biophys. Acta

    (1998)
  • M. Komori et al.

    Biochim. Biophys. Acta

    (1992)
  • M. Komori et al.

    Biochim. Biophys. Acta

    (1992)
  • T. Igarashi et al.

    Arch. Biochem. Biophys.

    (1997)
  • T. Sakuma et al.

    Biochem. Pharmacol.

    (1998)
  • E. Jacqz-Aigrain et al.

    Biochem. Pharmacol.

    (1991)
  • F.P. Guengerich

    J. Biol. Chem.

    (1977)
  • T. Omura et al.

    J. Biol. Chem.

    (1964)
  • T. Kronbach et al.

    Anal. Biochem.

    (1987)
  • T. Prueksaritanont et al.

    Biochem. Pharmacol.

    (1995)
  • M.S. Ching et al.

    Biochem. Pharmacol.

    (1995)
  • O. Gotoh

    J. Biol. Chem.

    (1992)
  • D.R. Nelson et al.

    Pharmacogenetics

    (1996)
  • P. Soucek et al.

    Xenobiotica

    (1992)
  • S.A. Wrighton et al.

    Crit. Rev. Toxicol.

    (1992)
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    The nucleotide sequence encoding CYP2D17 appears in GenBank under Accession No. U38218, and the subfamily designation was assigned by David Nelson of the CYP Nomenclature Committee.

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    Present address: Lilly Research Labs, Lilly Corporate Center, Indianapolis, IN 46285.

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    Present address: Bristol Myers Squibb, 5 Research Parkway, Wallingford, CT 06942.

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