2006
Study of recombinant cytochrome P450 2C9 activity with diclofenac by micellar electrokinetic capillary chromatography
KONEČNÝ, Jiří a Zdeněk GLATZZákladní údaje
Originální název
Study of recombinant cytochrome P450 2C9 activity with diclofenac by micellar electrokinetic capillary chromatography
Název česky
Studií aktivity rekombinantní izoformy cytochromu P450 2C9 pomocí MEKC
Autoři
KONEČNÝ, Jiří a Zdeněk GLATZ
Vydání
Belgium, CD of the abstracts of the 29th international symposium on capillary chromatography, od s. x, 1 s. 2006
Nakladatel
T. Sandra and P. Sandra
Další údaje
Jazyk
angličtina
Typ výsledku
Stať ve sborníku
Obor
10600 1.6 Biological sciences
Stát vydavatele
Itálie
Utajení
není předmětem státního či obchodního tajemství
Organizační jednotka
Přírodovědecká fakulta
Klíčová slova anglicky
cytochromes P450; diclofenac; MEKC
Štítky
Změněno: 19. 5. 2009 18:44, prof. RNDr. Zdeněk Glatz, CSc.
V originále
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.
Česky
Cíem práce bylo vypracovat MEKC metodu pro sledování aktivity izoformy cytochromu P450 2C9 pomocí MEKC.
Návaznosti
| GA203/06/0047, projekt VaV |
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| LC06023, projekt VaV |
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| MSM0021622413, záměr |
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