Study of Enzyme Kinetics of Phenol Sulfotransferase using EMMA

D 2003

Study of Enzyme Kinetics of Phenol Sulfotransferase using EMMA

NOVÁKOVÁ, Soňa, Sigrid VAN DYCK, Ann VAN SCHEPDAEL, Jos HOGMARTENS, Zdeněk GLATZ et. al.

Basic information

Original name

Study of Enzyme Kinetics of Phenol Sulfotransferase using EMMA

Authors

NOVÁKOVÁ, Soňa (203 Czech Republic), Sigrid VAN DYCK (56 Belgium), Ann VAN SCHEPDAEL (56 Belgium), Jos HOGMARTENS (56 Belgium) and Zdeněk GLATZ (203 Czech Republic, guarantor)

Edition

Nice, France, Proceeding of 27th Symposium on HPLC and Related Techniques, p. 133-133, 1 pp. 2003

Publisher

MCI Paris France

Other information

Language

English

Type of outcome

Stať ve sborníku

Field of Study

10600 1.6 Biological sciences

Country of publisher

France

Confidentiality degree

není předmětem státního či obchodního tajemství

RIV identification code

RIV/00216224:14310/03:00008126

Organization unit

Faculty of Science

Keywords in English

CZE; Enzymes; Phenol Sulfotransferase

Abstract

V originále

Sulfonation represents a major pathway for the biotransformation/detoxication of drugs and xenobiotics, as well as endogenous compounds such as cholesterol, catecholamines, steroid and thyroid hormones, and bile acid. The responsible enzymes are called the cytosolic sulfotransferases. This family of sulfotransferases (SULT) belongs to the Phase II group of drug metabolizing enzymes and catalyzes the sulfonation reaction involving the transfer of an electrophilic sulfonate group from the co-substrate adenosine 3-phosphate 5-phosphosulfate (PAPS) to a nucleophilic acceptor substrate. Humans have at least 11 different SULT enzymes, forming three subfamilies based on their amino acid sequence identity and substrate specifity: SULT1 (phenol sulfotransferase family), SULT2 (hydroxysteroid sulfotransferase family), SULT4 (brain sulfotransferase). In this work we used an isoenzyme from the SULT1A subfamily that is responsible for the sulfonation of small phenolic substrates such as p-nitrophenol, 2-naphtol and dopamine. The purpose of this study was to determine whether it is possible to assay SULT1A (EC 2.8.2.-) activity with p-nitrophenol and dopamine as different substrates by capillary electrophoresis. Capillary electrophoretic systems have been successfully applied for on-line enzyme-catalyzed reactions by a methodology known as electrophoretically mediated microanalysis (EMMA), firstly described by Bao and Regnier. EMMA utilizes the different electrophoretic mobilities of enzyme, substrate, and product to initiate reaction inside the capillary and to separate the components from each other for final in-capillary quantitation. Compared to spectrophotometric and other discontinuous assays, the method is rapid, can be automated, does not need expensive radiolabeled substrates and requires only small amounts of reagents, which is especially important in the case of enzymes (injection volumes of analytes are in the order of nl). In consequence the EMMA methodology could serve as a progressive tool of modern enzymology in the context of metabolic research.

Links

GA203/03/1125, research and development project

Name: Využití kapilární zónové elektroforézy pro studium enzymů

Investor: Czech Science Foundation, Application of capillary zone electrophoresis for the study of enzymes

Citovat

NOVÁKOVÁ, Soňa, Sigrid VAN DYCK, Ann VAN SCHEPDAEL, Jos HOGMARTENS and Zdeněk GLATZ. Study of Enzyme Kinetics of Phenol Sulfotransferase using EMMA. In Proceeding of 27th Symposium on HPLC and Related Techniques. Nice, France: MCI Paris France, 2003, p. 133-133.

@inproceedings{488017,
   author = {Nováková, Soňa and Van Dyck, Sigrid and Van Schepdael, Ann and Hogmartens, Jos and Glatz, Zdeněk},
   address = {Nice, France},
   booktitle = {Proceeding of 27th Symposium on HPLC and Related Techniques},
   keywords = {CZE; Enzymes; Phenol Sulfotransferase},
   language = {eng},
   location = {Nice, France},
   pages = {133-133},
   publisher = {MCI Paris France},
   title = {Study of Enzyme Kinetics of Phenol Sulfotransferase using EMMA},
   year = {2003}
}

TY  - JOUR
ID  - 488017
AU  - Nováková, Soňa - Van Dyck, Sigrid - Van Schepdael, Ann - Hogmartens, Jos - Glatz, Zdeněk
PY  - 2003
TI  - Study of Enzyme Kinetics of Phenol Sulfotransferase using EMMA
PB  - MCI Paris France
CY  - Nice, France
KW  - CZE
KW  - Enzymes
KW  - Phenol Sulfotransferase
N2  - Sulfonation represents a major pathway for the biotransformation/detoxication of drugs and xenobiotics, as well as endogenous compounds such as cholesterol, catecholamines, steroid and thyroid hormones, and bile acid. The responsible enzymes are called the cytosolic sulfotransferases. This family of sulfotransferases (SULT) belongs to the Phase II group of drug metabolizing enzymes and catalyzes the sulfonation reaction involving the transfer of an electrophilic sulfonate group from the co-substrate adenosine 3-phosphate 5-phosphosulfate (PAPS) to a nucleophilic acceptor substrate. Humans have at least 11 different SULT enzymes, forming three subfamilies based on their amino acid sequence identity and substrate specifity: SULT1 (phenol sulfotransferase family), SULT2 (hydroxysteroid sulfotransferase family), SULT4 (brain sulfotransferase). In this work we used an isoenzyme from the SULT1A subfamily that is responsible for the sulfonation of small phenolic substrates such as p-nitrophenol, 2-naphtol and dopamine. The purpose of this study was to determine whether it is possible to assay SULT1A (EC 2.8.2.-) activity with p-nitrophenol and dopamine as different substrates by capillary electrophoresis. Capillary electrophoretic systems have been successfully applied for on-line enzyme-catalyzed reactions by a methodology known as electrophoretically mediated microanalysis (EMMA), firstly described by Bao and Regnier. EMMA utilizes the different electrophoretic mobilities of enzyme, substrate, and product to initiate reaction inside the capillary and to separate the components from each other for final in-capillary quantitation. Compared to spectrophotometric and other discontinuous assays, the method is rapid, can be automated, does not need expensive radiolabeled substrates and requires only small amounts of reagents, which is especially important in the case of enzymes (injection volumes of analytes are in the order of nl). In consequence the EMMA methodology could serve as a progressive tool of modern enzymology in the context of metabolic research.
ER  -

NOVÁKOVÁ, Soňa, Sigrid VAN DYCK, Ann VAN SCHEPDAEL, Jos HOGMARTENS and Zdeněk GLATZ. Study of Enzyme Kinetics of Phenol Sulfotransferase using EMMA. In \textit{Proceeding of 27th Symposium on HPLC and Related Techniques}. Nice, France: MCI Paris France, 2003, p.~133-133.

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