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A fluorometric method for determination of catalytic activity of CYP51b1 (sterol 14α-demethylase) with coumarin derivatives

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Abstract

A method for direct registration of CYP51b1 (sterol-14α-demethylase) activity with coumarin derivatives as substrate has been proposed. Determination of catalytic activity of this enzyme with 7-aminocoumarin-4-acetic acid (ACAC) is based on registration of the increase of fluorescence (λexcitation = 360 nm and λemission = 435 nm) at 30°C. In the model system for assay of CYP51b1 activity BMR (a flavin domain of the bacterial cytochrome P450BM-3) may serve as the electron donor. The developed method is simple, highly sensitive and accurate; it can be used for screening of sterol 14α-demethylase inhibitors.

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Authors and Affiliations

  1. Institute of Biomedical Chemistry, Russian Academy of Medical Sciences, ul Pogodinskaya 10, Moscow, 119121, Russia

    N. A. Petushkova, A. V. Lisitsa, V. F. Pozdnev & I. I. Karuzina

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  1. N. A. Petushkova
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  2. A. V. Lisitsa
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  3. V. F. Pozdnev
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  4. I. I. Karuzina
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Correspondence to N. A. Petushkova.

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Original Russian Text © N.A. Petushkova, A.V. Lisitsa, V.F. Pozdnev, I.I. Karuzin, 2010, published in Biomeditsinskaya Khimiya.

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Petushkova, N.A., Lisitsa, A.V., Pozdnev, V.F. et al. A fluorometric method for determination of catalytic activity of CYP51b1 (sterol 14α-demethylase) with coumarin derivatives. Biochem. Moscow Suppl. Ser. B 4, 104–106 (2010). https://doi.org/10.1134/S1990750810010130

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  • DOI: https://doi.org/10.1134/S1990750810010130

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