Literature DB >> 10574186

Novel sensitive high-performance liquid chromatographic method for assay of coumarin 7-hydroxylation.

P Soucek1.   

Abstract

In this paper, a novel HPLC-based method with fluorometric detection of coumarin 7-hydroxylase is presented. The described method provides a time-effective, more sensitive and specific alternative to the previously used spectrofluorometric assay. Using the developed method, metabolism of coumarin in 11 samples of human liver microsomes was evaluated and 1790+/-690 pmol/min/nmol cytochrome P450 (CYP) activity was found. Kinetic parameters and linearity of coumarin 7-hydroxylation were studied in a reconstituted system consisting of recombinant CYP2A6 expressed in Escherichia coli, rat NADPH-CYP reductase and usual components. It was found that a 3.5 to 30 min time of incubation is suitable for estimation of coumarin 7-hydroxylase activity. Observed Km and Vmax values in the CYP2A6 reconstituted system were 1.48+/-0.37 microM and 3360+/-180 pmol product/min/nmol CYP, respectively.

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Year:  1999        PMID： 10574186

Source DB:  PubMed          Journal:  J Chromatogr B Biomed Sci Appl        ISSN： 1387-2273


  7 in total

1.  Mechanism-based inhibition of cytochrome P450 (CYP)2A6 by chalepensin in recombinant systems, in human liver microsomes and in mice in vivo.

Authors:  Yune-Fang Ueng; Chien-Chih Chen; Yu-Ting Chung; Tsung-Yun Liu; Yu-Ping Chang; Wei-Sheng Lo; Norie Murayama; Hiroshi Yamazaki; Pavel Souček; Gar-Yang Chau; Chin-Wen Chi; Ruei-Ming Chen; Ding-Tzai Li
Journal:  Br J Pharmacol       Date:  2011-07       Impact factor: 8.739

2.  In vitro and in silico studies of interaction of synthetic 2,6,9-trisubstituted purine kinase inhibitors BPA-302, BP-21 and BP-117 with liver drug-metabolizing cytochromes P450

Authors:  A Špičáková; P Kraus; T Gucký; V Kryštof; M Strnad; V Bazgier; M Otyepka; V Kubíčková; M Poruba; Z Rácová; I Zapletalová; P Anzenbacher
Journal:  Physiol Res       Date:  2020-12-31       Impact factor: 1.881

3.  Minipig cytochrome P450 3A, 2A and 2C enzymes have similar properties to human analogs.

Authors:  P Soucek; R Zuber; E Anzenbacherová; P Anzenbacher; F P Guengerich
Journal:  BMC Pharmacol       Date:  2001-12-05

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Authors:  Alena Špičáková; Barbora Szotáková; Diana Dimunová; Zuzana Myslivečková; Vladimír Kubíček; Martin Ambrož; Kateřina Lněničková; Kristýna Krasulová; Pavel Anzenbacher; Lenka Skálová
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5.  Transcriptional and Epigenetic Consequences of DMSO Treatment on HepaRG Cells.

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Journal:  Cells       Date:  2022-07-26       Impact factor: 7.666

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Authors:  Yune-Fang Ueng; Chien-Chih Chen; Yu-Ling Huang; I-Jung Lee; Chul-Ho Yun; Yu-Hsuan Chen; Chiung-Chiao Huang
Journal:  J Food Drug Anal       Date:  2015-05-21       Impact factor: 6.157

7.  The time-dependent effects of St John's wort on cytochrome P450, uridine diphosphate-glucuronosyltransferase, glutathione S-transferase, and NAD(P)H-quinone oxidoreductase in mice.

Authors:  Jin-Fu Yang; Yue-Rong Liu; Chiung-Chiao Huang; Yune-Fang Ueng
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  7 in total

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