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@article{492938,
author = {Vytisková, Soňa and Van Dyck, Sigrid and Van Schepdael, Ann and Hoogmartens, Jos and Glatz, Zdeněk},
article_location = {Netherlands},
article_number = {(1+2)},
keywords = {Capillary electrophoresis; enzymes; EMMA; sulfotranferase},
language = {eng},
issn = {0021-9606},
journal = {Journal of Chromatography A},
title = {Study of enzyme kinetics of phenol sulfotransferase by electrophoretically mediated microanalysis},
volume = {1032},
year = {2004}
}
TY - JOUR
ID - 492938
AU - Vytisková, Soňa - Van Dyck, Sigrid - Van Schepdael, Ann - Hoogmartens, Jos - Glatz, Zdeněk
PY - 2004
TI - Study of enzyme kinetics of phenol sulfotransferase by electrophoretically mediated microanalysis
JF - Journal of Chromatography A
VL - 1032
IS - (1+2)
SP - 319-326
EP - 319-326
PB - Elsevier
SN - 00219606
KW - Capillary electrophoresis
KW - enzymes
KW - EMMA
KW - sulfotranferase
N2 - Electrophoretically mediated microanalysis (EMMA) was applied for the study of the kinetic parameters of the enzymatic reaction of phenol sulfotransferase SULT1A1 isoenzyme with 4-nitrophenol as a substrate. The SULT1A1 activity was determined by the quantitation of the product, 4-nitrophenyl sulfate, at 274 nm by using different injection and separation steps. This new approach solved the problem of the presence of the very strong inhibitor, adenosine 3',5'-bisphosphate (PAP), in the co-substrate solution (adenosine 3'-phosphate 5'-phosphosulfate, PAPS) which is unstable at room temperature. The inhibitor PAP was electrophoretically separated from the co-substrate PAPS before the injection of enzyme and substrate inside the capillary (and thus before their in-capillary encountering). With the developed in-capillary SULT1A1 activity assay an average Michaelis constant (Km) for 4-nitrophenol was calculated to be 0.84 M, a value which is consistent with a previously reported value. Strong substrate inhibition (above a 4-nitrophenol concentration of 2.5 M) was observed, and this is also in accordance with literature values.
ER -
VYTISKOVÁ, Soňa, Sigrid VAN DYCK, Ann VAN SCHEPDAEL, Jos HOOGMARTENS a Zdeněk GLATZ. Study of enzyme kinetics of phenol sulfotransferase by electrophoretically mediated microanalysis. \textit{Journal of Chromatography A}. Netherlands: Elsevier, 2004, roč.~1032, (1+2), s.~319-326. ISSN~0021-9606.
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