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Description of a 96-Well Plate Assay to Measure Cytochrome P4503A Inhibition in Human Liver Microsomes Using a Selective Fluorescent Probe by Nathalie Chauret; Nathalie Tremblay; Rebecca L. Lackman; Jacques-Yves Gauthier; José M. Silva; Jean Marois; James A. Yergey; Deborah A. Nicoll-Griffith is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.
What is Description of a 96-Well Plate Assay to Measure Cytochrome P4503A Inhibition in Human Liver Microsomes Using a Selective Fluorescent Probe about?
The standard method to evaluate CYP3A inhibition is to study the conversion of the specific CYP3A probe testosterone to its 6-hydroxy metabolite in human liver microsomes, in the absence and presence of potential inhibitors. Quantification of the 6-hydroxy metabolite is achieved by HPLC resulting in a tedious and time-consuming assay. In order to increase the P450 inhibition throughput, efforts were made to find a CYP3A probe that would produce a fluorescent metabolite. This paper reports the discovery of DFB as a potential CYP3A fluorescent probe. DFB was significantly metabolized in human microsomes (ϳ1-2 nmol/ (min 1⁄7 mg protein)) to give the fluorescent compound DFH. The involvement of CYP3A in the metabolism of DFB was determined using multiple approaches. First, incubations conducted with microsomes made from cell lines expressing single CYPs (Gentest Supersomes) indicated that CYP3A played a major role in the metabolism of DFB. Secondly, immunoinhibition studies conducted with CYP3A antibody resulted in >95% inhibition of DFB metabolism in HLM. Thirdly, inhibition studies with specific CYP1A1, 1A2, 2C8/9, 2C19, 2D6, and 2E1 chemical inhibitors did not suppress DFB activit
Who reads Description of a 96-Well Plate Assay to Measure Cytochrome P4503A Inhibition in Human Liver Microsomes Using a Selective Fluorescent Probe?
It is typically read by researchers, students, and practitioners in Biochemistry, Genetics and Molecular Biology.
- Author
- Nathalie Chauret; Nathalie Tremblay; Rebecca L. Lackman; Jacques-Yves Gauthier; José M. Silva; Jean Marois; James A. Yergey; Deborah A. Nicoll-Griffith
- Publisher
- Elsevier Science; Elsevier ; Elsevier Inc.; Elsevier BV (ISSN 0003-2697)
- Published
- 1999
- Language
- EN
- Field
- Biochemistry, Genetics and Molecular Biology (Life Sciences)