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Rapid CommunicationShort Communication

Expression and Characterization of Dog Cytochrome P450 2A13 and 2A25 in Baculovirus-Infected Insect Cells

Diansong Zhou, Alban J. Linnenbach, Ruifeng Liu, Rick A. Luzietti, Jennifer J. Harris, Catherine L. Booth-Genthe and Scott W. Grimm
Drug Metabolism and Disposition July 2010, 38 (7) 1015-1018; DOI: https://doi.org/10.1124/dmd.110.033068
Diansong Zhou
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Alban J. Linnenbach
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Ruifeng Liu
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Rick A. Luzietti
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Jennifer J. Harris
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Catherine L. Booth-Genthe
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Scott W. Grimm
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Abstract

Dog CYP2A13 and CYP2A25 were coexpressed with dog NADPH-cytochrome P450 reductase (OR) in baculovirus-infected Sf9 insect cells. CYP2A13 effectively catalyzed 7-ethoxycoumarin (7EC) deethylation and coumarin hydroxylation with apparent Km values of 4.8 and 2.1 μM, respectively, similar to those observed using dog liver microsomes (7.5 and 0.75 μM, respectively). CYP2A25 exhibited much lower affinity toward 7EC, with an apparent Km value of 150 μM, which indicates that CYP2A13 plays a more significant role in the metabolism of these CYP2A substrates. Similar to the dog CYP1A2 enzyme, CYP2A13 efficiently catalyzed phenacetin deethylation with a Km value of 3.9 μM, which suggests that phenacetin is not a selective probe for dog CYP1A2 activity. Both dog CYP2A13 and CYP2A25 exhibited little or no catalytic activity toward other common cytochrome P450 probe substrates, including bupropion, amodiaquine, diclofenac, S-mephenytoin, bufuralol, dextromethorphan, midazolam, and testosterone. These results provided additional information about the selectivity of these commonly used probe substrates.

Footnotes

  • Article, publication date, and citation information can be found at http://dmd.aspetjournals.org.

    doi:10.1124/dmd.110.033068.

  • ABBREVIATIONS:

    P450
    cytochrome P450
    OR
    NADPH-cytochrome P450 reductase
    PCR
    polymerase chain reaction
    7EC
    7-ethoxycoumarin.

  • Received March 2, 2010.
  • Accepted April 9, 2010.
  • Copyright © 2010 by The American Society for Pharmacology and Experimental Therapeutics
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Drug Metabolism and Disposition: 38 (7)
Drug Metabolism and Disposition
Vol. 38, Issue 7
1 Jul 2010
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Rapid CommunicationShort Communication

Expression and Characterization of Dog Cytochrome P450 2A13 and 2A25 in Baculovirus-Infected Insect Cells

Diansong Zhou, Alban J. Linnenbach, Ruifeng Liu, Rick A. Luzietti, Jennifer J. Harris, Catherine L. Booth-Genthe and Scott W. Grimm
Drug Metabolism and Disposition July 1, 2010, 38 (7) 1015-1018; DOI: https://doi.org/10.1124/dmd.110.033068

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Rapid CommunicationShort Communication

Expression and Characterization of Dog Cytochrome P450 2A13 and 2A25 in Baculovirus-Infected Insect Cells

Diansong Zhou, Alban J. Linnenbach, Ruifeng Liu, Rick A. Luzietti, Jennifer J. Harris, Catherine L. Booth-Genthe and Scott W. Grimm
Drug Metabolism and Disposition July 1, 2010, 38 (7) 1015-1018; DOI: https://doi.org/10.1124/dmd.110.033068
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