KONEČNÝ, Jiří and Zdeněk GLATZ. Study of recombinant cytochrome P450 2C9 activity with diclofenac by micellar electrokinetic capillary chromatography. In CD of the abstracts of the 29th international symposium on capillary chromatography. Belgium: T. Sandra and P. Sandra, 2006, p. x, 1 pp.
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Basic information
Original name Study of recombinant cytochrome P450 2C9 activity with diclofenac by micellar electrokinetic capillary chromatography
Name in Czech Studií aktivity rekombinantní izoformy cytochromu P450 2C9 pomocí MEKC
Authors KONEČNÝ, Jiří and Zdeněk GLATZ.
Edition Belgium, CD of the abstracts of the 29th international symposium on capillary chromatography, p. x, 1 pp. 2006.
Publisher T. Sandra and P. Sandra
Other information
Original language English
Type of outcome Proceedings paper
Field of Study 10600 1.6 Biological sciences
Country of publisher Italy
Confidentiality degree is not subject to a state or trade secret
Organization unit Faculty of Science
Keywords in English cytochromes P450; diclofenac; MEKC
Tags cytochromes P450, diclofenac, MEKC
Changed by Changed by: prof. RNDr. Zdeněk Glatz, CSc., učo 1865. Changed: 19/5/2009 18:44.
Abstract
The cytochromes P450 [EC 1.14.14.1] comprise a family of b-type heme containing proteins with an identical prosthetic group and mechanism of catalysis, which are responsible for the biotransformation of xenobiotics and endogenous compounds. Because of the pharmacological relevance of this enzyme different analytical methods have been developed to measure the expression of cytochrome P450 in biological samples. The most common methods are determination of the catalytic activities of individual cytochrome P450 isoformes using model substrates. The purpose of this communication is to demonstrate the applicability of capillary electrophoresis for the assay of the enzymatic activity of the human cytochrome P450 2C9 (CYP 2C9), one of the most important isoform in human liver involved in the metabolism of a large number of therapeutic agents. Micellar electrokinetic capillary chromatography (MEKC) with SDS as pseudostationary phase was used for this purpose with diclofenac as a substrate. The method was also applied to evaluate some properties of the CYP 2C9 including Michaelis constant, pH and temperature optima of enzymatic reaction. Compared to other assays, the MEKC based method is rapid, can be automated, and requires only small quantity of enzyme and substrate. Moreover, the enzymatic reaction can be monitored with high sensitivity and reproducibility even when the reaction mixture without any pre-treatment is used for the analysis.
Abstract (in Czech)
Cíem práce bylo vypracovat MEKC metodu pro sledování aktivity izoformy cytochromu P450 2C9 pomocí MEKC.
Links
GA203/06/0047, research and development projectName: Využití kapilární elektroforézy pro studium metabolismu léčiv
Investor: Czech Science Foundation, Capillary electrophoresis as a tool for the drug-metabolism studies
LC06023, research and development projectName: Integrované bioanalytické technologie pro mikroanalýzy a diagnostiku s využitím LIF a hmotnostní spektrometrie
Investor: Ministry of Education, Youth and Sports of the CR
MSM0021622413, plan (intention)Name: Proteiny v metabolismu a při interakci organismů s prostředím
Investor: Ministry of Education, Youth and Sports of the CR, Proteins in metabolism and interaction of organisms with the environment
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