Study of cytosolic glutathione s-tranferase activity with
styrene oxide by electromigration methods

D 2004

Study of cytosolic glutathione s-tranferase activity with styrene oxide by electromigration methods

ŠIŠKOVÁ, Zdeňka, Soňa VYTISKOVÁ, Zdeněk GLATZ and Vladislav KAHLE

Basic information

Original name

Study of cytosolic glutathione s-tranferase activity with styrene oxide by electromigration methods

Name in Czech

Studium cytosolické GST pomocí elektromigračních metod.

Authors

ŠIŠKOVÁ, Zdeňka (203 Czech Republic), Soňa VYTISKOVÁ (203 Czech Republic), Zdeněk GLATZ (203 Czech Republic, guarantor) and Vladislav KAHLE (203 Czech Republic)

Edition

Řím, Book of abstract 14 th International Symposium on Capillary Electroseparation Techniques ITP 2004, p. 32-32, 2004

Publisher

Consiglio Nazionale delle Ricerche

Other information

Language

English

Type of outcome

Stať ve sborníku

Field of Study

10600 1.6 Biological sciences

Country of publisher

Italy

Confidentiality degree

není předmětem státního či obchodního tajemství

RIV identification code

RIV/00216224:14310/04:00010320

Organization unit

Faculty of Science

Keywords in English

GST; capillary electrochromatography; MEKC

Abstract

ORIG CZ

V originále

Styrene-7,8-oxide is an industrial chemical used in the manufacture of epoxy resins; as an intermediate in the preparation of various agricultural chemicals, cosmetics and plastics; and in the processing of textile and fibres. SO is also the major cytochrome P450 metabolite of styrene, used in plastic and resin manufacture. As epoxide, SO is highly reactive and able to undergo reactions with various nucleophilic groups in tissue components, causing radiomimetic effects such as growth inhibition, cytotoxicity and mutagenicity. In addition SO has been classified as carcinogenic in animals and is probably carcinogenic in humans. Its detoxification may occur due to glutathione S-transferase (GST) catalysed conjugation with cytosolic glutathione. As people working in a number of industries are exposed to styrene and SO, it is important to examine the role of glutathione conjugation versus other competing detoxication pathways. Similar studies have often been hampered by the lack of reliable methods for quantitating the glutathione adducts. In this study two electromigration methods - capillary electrochromatography (CEC) in silica-based polymeric monolithic columns and micellar electrokinetic capillary chromatography (MEKC) with SDS as pseudostationary phase were tested for this purpose. Because of relatively hydrophilic nature of given conjugates no separation was achieved by CEC whereas good separation of conjugates was obtained by MEKC. The method based on MEKC was further optimised and used for measuring GST activity with SO. Unlike previous methods, proposed assay did not require the use of radiolabelled substrates, and was sufficiently sensitive to detect conjugate formation in tissues with low GST activity. In addition it could be applied for determination of kinetic parameters of GST - pH and temperature optima of enzymatic reaction, and Michaelis constants (Km) for SO.

In Czech

jedná se o studium cytosolické GST pomocí elektromigračních metod.

Links

GA203/02/1447, research and development project

Name: Kapilární elektrochromatografie v lineárních a planárních mikrostrukturách s využitím polymerních náplní

Investor: Czech Science Foundation, Capillary electrochromatography in linear and planar microstructures using polymer beds

MSM 143100005, plan (intention)

Name: Strukturně-funkční vztahy biomolekul a jejich role v metabolismu

Investor: Ministry of Education, Youth and Sports of the CR, Biomolecular Structure-function Relationships and their role in the Metabolism

Citovat

ŠIŠKOVÁ, Zdeňka, Soňa VYTISKOVÁ, Zdeněk GLATZ and Vladislav KAHLE. Study of cytosolic glutathione s-tranferase activity with styrene oxide by electromigration methods. In Book of abstract 14 th International Symposium on Capillary Electroseparation Techniques ITP 2004. Řím: Consiglio Nazionale delle Ricerche, 2004, p. 32.

@inproceedings{558258,
   author = {Šišková, Zdeňka and Vytisková, Soňa and Glatz, Zdeněk and Kahle, Vladislav},
   address = {Řím},
   booktitle = {Book of abstract 14 th International Symposium on Capillary Electroseparation Techniques ITP 2004},
   keywords = {GST; capillary electrochromatography; MEKC},
   language = {eng},
   location = {Řím},
   pages = {32-32},
   publisher = {Consiglio Nazionale delle Ricerche},
   title = {Study of cytosolic glutathione s-tranferase activity with styrene oxide by electromigration methods},
   year = {2004}
}

TY  - JOUR
ID  - 558258
AU  - Šišková, Zdeňka - Vytisková, Soňa - Glatz, Zdeněk - Kahle, Vladislav
PY  - 2004
TI  - Study of cytosolic glutathione s-tranferase activity with styrene oxide by electromigration methods
PB  - Consiglio Nazionale delle Ricerche
CY  - Řím
KW  - GST
KW  - capillary electrochromatography
KW  - MEKC
N2  - Styrene-7,8-oxide is an industrial chemical used in the manufacture of epoxy resins; as an intermediate in the preparation of various agricultural chemicals, cosmetics and plastics; and in the processing of textile and fibres. SO is also the major cytochrome P450 metabolite of styrene, used in plastic and resin manufacture. As epoxide, SO is highly reactive and able to undergo reactions with various nucleophilic groups in tissue components, causing radiomimetic effects such as growth inhibition, cytotoxicity and mutagenicity. In addition SO has been classified as carcinogenic in animals and is probably carcinogenic in humans. Its detoxification may occur due to glutathione S-transferase (GST) catalysed conjugation with cytosolic glutathione. As people working in a number of industries are exposed to styrene and SO, it is important to examine the role of glutathione conjugation versus other competing detoxication pathways. Similar studies have often been hampered by the lack of reliable methods for quantitating the glutathione adducts. In this study two electromigration methods - capillary electrochromatography (CEC) in silica-based polymeric monolithic columns and micellar electrokinetic capillary chromatography (MEKC) with SDS as pseudostationary phase were tested for this purpose. Because of relatively hydrophilic nature of given conjugates no separation was achieved by CEC whereas good separation of conjugates was obtained by MEKC. The method based on MEKC was further optimised and used for measuring GST activity with SO. Unlike previous methods, proposed assay did not require the use of radiolabelled substrates, and was sufficiently sensitive to detect conjugate formation in tissues with low GST activity. In addition it could be applied for determination of kinetic parameters of GST - pH and temperature optima of enzymatic reaction, and Michaelis constants (Km) for SO.
ER  -

ŠIŠKOVÁ, Zdeňka, Soňa VYTISKOVÁ, Zdeněk GLATZ and Vladislav KAHLE. Study of cytosolic glutathione s-tranferase activity with styrene oxide by electromigration methods. In \textit{Book of abstract 14 th International Symposium on Capillary Electroseparation Techniques ITP 2004}. Řím: Consiglio Nazionale delle Ricerche, 2004, p.~32.

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