Fluorescence-based determination of enzyme activity of recombinant CYP51b1 (sterol 14α-demethylase) with coumarin derivatives

   


1. Institute of Biomedical Chemistry, Russian Academy of Medical Sciences
Type: Experimental/clinical study
DOI: 10.18097/pbmc20105601132      UDK: 577.15+612.853-205      PubMed Id: 21328917
Year: 2010 vol: 56  issue:1  pages: 132-137
Abstract: The current investigation was undertaken with the aim to carry out an in vitro evaluation of the ability of coumarin derivatives as probe substrates to predict the activity of CYP51b1. The results obtained indicate that 7-aminocoumarin-4-acetic acid (ACAC) can be used to determine the recombinant CYP51b1 activity. Determination of CYP51b1 activity with ACAC is based on the direct registration of fluorescence increasing at 30°C. It was found also that BMR in a simple soluble model system can be used as an electron donor for CYP51B1. Fluorescence-based assay is highly sensitive and can be used for the screening of sterol 14alpha-demethylase inhibitors.
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Reference: Petushkova N.A., Lisitsa A.V., Pozdnev V.F., Karuzina I.I., Fluorescence-based determination of enzyme activity of recombinant CYP51b1 (sterol 14α-demethylase) with coumarin derivatives, Biomeditsinskaya khimiya, 2010, vol: 56(1), 132-137.
This paper is also available as the English translation:10.1134/S1990750810010130
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