High-Throughput Functional Screening of Steroid Substrates with Wild-Type and Chimeric P450 Enzymes
Joint Authors
Urban, Philippe
Truan, Gilles
Pompon, Denis
Source
Issue
Vol. 2014, Issue 2014 (31 Dec. 2014), pp.1-11, 11 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2014-08-26
Country of Publication
Egypt
No. of Pages
11
Main Subjects
Abstract EN
The promiscuity of a collection of enzymes consisting of 31 wild-type and synthetic variants of CYP1A enzymes was evaluated using a series of 14 steroids and 2 steroid-like chemicals, namely, nootkatone, a terpenoid, and mifepristone, a drug.
For each enzyme-substrate couple, the initial steady-state velocity of metabolite formation was determined at a substrate saturating concentration.
For that, a high-throughput approach was designed involving automatized incubations in 96-well microplate with sixteen 6-point kinetics per microplate and data acquisition using LC/MS system accepting 96-well microplate for injections.
The resulting dataset was used for multivariate statistics aimed at sorting out the correlations existing between tested enzyme variants and ability to metabolize steroid substrates.
Functional classifications of both CYP1A enzyme variants and steroid substrate structures were obtained allowing the delineation of global structural features for both substrate recognition and regioselectivity of oxidation.
American Psychological Association (APA)
Urban, Philippe& Truan, Gilles& Pompon, Denis. 2014. High-Throughput Functional Screening of Steroid Substrates with Wild-Type and Chimeric P450 Enzymes. BioMed Research International،Vol. 2014, no. 2014, pp.1-11.
https://search.emarefa.net/detail/BIM-1016546
Modern Language Association (MLA)
Urban, Philippe…[et al.]. High-Throughput Functional Screening of Steroid Substrates with Wild-Type and Chimeric P450 Enzymes. BioMed Research International No. 2014 (2014), pp.1-11.
https://search.emarefa.net/detail/BIM-1016546
American Medical Association (AMA)
Urban, Philippe& Truan, Gilles& Pompon, Denis. High-Throughput Functional Screening of Steroid Substrates with Wild-Type and Chimeric P450 Enzymes. BioMed Research International. 2014. Vol. 2014, no. 2014, pp.1-11.
https://search.emarefa.net/detail/BIM-1016546
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-1016546