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A Microassay for the detection of low levels of cytochrome P450 O-deethylation activities with alkoxyresorufin substrates

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Abstract

A microfluorometric method for the detection of low levels of cytochrome P450 was developed to increase the sensitivity of the assay, since a low level of CYP450 associated enzymatic activities was expected in human placenta tissues and small samples of placenta (∼10 g) could be easily collected, stored and processed. The dual fluorescence assay of Kennedy et al. [1], which was developed to simultaneously quantitate microsomal proteins and ethoxyresorufin-O-deethylase (EROD) activity was adapted for 96 wells microtiter plates. Placental microsomes samples were analyzed. For samples obtained from non-smoking mothers from the general southern Quebec population, results ranged from less than 1–3.3 pmol/mg protein•min. Samples collected from smoking mothers showed activity levels ranging from 30–69 pmol/mg protein•min. These results showed the suitability of the microassay for measuring low level of CYP450 activity in tissues such as placenta. (Mol Cell Biochem 175: 131–136, 1997)

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Authors and Affiliations

  1. Health and Environment Unit, Centre Hospitalier Universitaire de Québec Research Center, Ste-Foy (, Qc), Canada

    Jean Lagueux, Denis Nadeau, Pierre Ayotte, Éric Dewailly & Guy G. Poirier

  2. Centre de Santé publique de Québec, Beauport (, Qc), Canada

    Éric Dewailly & Guy G. Poirier

Authors
  1. Jean Lagueux
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  2. Denis Nadeau
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  3. Pierre Ayotte
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  4. Éric Dewailly
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  5. Guy G. Poirier
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Lagueux, J., Nadeau, D., Ayotte, P. et al. A Microassay for the detection of low levels of cytochrome P450 O-deethylation activities with alkoxyresorufin substrates. Mol Cell Biochem 175, 125–129 (1997). https://doi.org/10.1023/A:1006835712436

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  • DOI: https://doi.org/10.1023/A:1006835712436

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