2004
Study of enzyme kinetics of phenol sulfotransferase by electrophoretically mediated microanalysis
VYTISKOVÁ, Soňa, Sigrid VAN DYCK, Ann VAN SCHEPDAEL, Jos HOOGMARTENS, Zdeněk GLATZ et. al.Základní údaje
Originální název
Study of enzyme kinetics of phenol sulfotransferase by electrophoretically mediated microanalysis
Název česky
Studium enzymove kinetiky enzymu fenolsulfotranferasy metodou EMMA
Autoři
VYTISKOVÁ, Soňa (203 Česká republika, garant), Sigrid VAN DYCK (56 Belgie), Ann VAN SCHEPDAEL (56 Belgie), Jos HOOGMARTENS (56 Belgie) a Zdeněk GLATZ (203 Česká republika)
Vydání
Journal of Chromatography A, Netherlands, Elsevier, 2004, 0021-9606
Další údaje
Jazyk
angličtina
Typ výsledku
Článek v odborném periodiku
Obor
10600 1.6 Biological sciences
Stát vydavatele
Nizozemské království
Utajení
není předmětem státního či obchodního tajemství
Impakt faktor
Impact factor: 3.105
Kód RIV
RIV/00216224:14310/04:00009928
Organizační jednotka
Přírodovědecká fakulta
UT WoS
000220121200043
Klíčová slova anglicky
Capillary electrophoresis; enzymes; EMMA; sulfotranferase
Štítky
Změněno: 19. 5. 2009 18:44, prof. RNDr. Zdeněk Glatz, CSc.
V originále
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.
Česky
Metoda EMMA byla využita pro studium enzymove kinetiky enzymu fenolsulfotranferasy.
Návaznosti
GA203/03/1125, projekt VaV |
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MSM 143100005, záměr |
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