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.

Základní údaje

Originální název

Study of Enzyme Kinetics of Phenol Sulfotransferase using EMMA

Autoři

NOVÁKOVÁ, Soňa (203 Česká republika), Sigrid VAN DYCK (56 Belgie), Ann VAN SCHEPDAEL (56 Belgie), Jos HOGMARTENS (56 Belgie) a Zdeněk GLATZ (203 Česká republika, garant)

Vydání

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

Nakladatel

MCI Paris France

Další údaje

Jazyk

angličtina

Typ výsledku

Stať ve sborníku

Obor

10600 1.6 Biological sciences

Stát vydavatele

Francie

Utajení

není předmětem státního či obchodního tajemství

Kód RIV

RIV/00216224:14310/03:00008126

Organizační jednotka

Přírodovědecká fakulta

Klíčová slova anglicky

CZE; Enzymes; Phenol Sulfotransferase
Změněno: 19. 5. 2009 18:44, prof. RNDr. Zdeněk Glatz, CSc.

Anotace

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.

Návaznosti

GA203/03/1125, projekt VaV
Název: Využití kapilární zónové elektroforézy pro studium enzymů
Investor: Grantová agentura ČR, Využití kapilární zónové elektroforézy pro studium enzymů