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 a Vladislav KAHLE

Základní údaje

Originální název

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

Název česky

Studium cytosolické GST pomocí elektromigračních metod.

Autoři

ŠIŠKOVÁ, Zdeňka (203 Česká republika), Soňa VYTISKOVÁ (203 Česká republika), Zdeněk GLATZ (203 Česká republika, garant) a Vladislav KAHLE (203 Česká republika)

Vydání

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

Nakladatel

Consiglio Nazionale delle Ricerche

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

Kód RIV

RIV/00216224:14310/04:00010320

Organizační jednotka

Přírodovědecká fakulta

Klíčová slova anglicky

GST; capillary electrochromatography; MEKC

Štítky

Capillary electrochromatography, GST, MEKC

Změněno: 19. 5. 2009 18:44, prof. RNDr. Zdeněk Glatz, CSc.

Anotace

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.

Česky

jedná se o studium cytosolické GST pomocí elektromigračních metod.

Návaznosti

GA203/02/1447, projekt VaV

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

Investor: Grantová agentura ČR, Kapilární elektrochromatografie v lineárních a planárních mikrostrukturách s využitím polymerních náplní

MSM 143100005, záměr

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

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, Strukturně-funkční vztahy biomolekul a jejich role v metabolismu

Citovat

ŠIŠKOVÁ, Zdeňka, Soňa VYTISKOVÁ, Zdeněk GLATZ a 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, s. 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 a 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, s.~32.

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