Open AccessIn‐vitro sulfation of piceatannol by human liver cytosol and recombinant sulfotransferases
Author(s) -
Miksits Michaela,
Sulyok Michael,
Schuhmacher Rainer,
Szekeres Thomas,
Jäger Walter
Publication year - 2009
Publication title -
journal of pharmacy and pharmacology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.745
H-Index - 118
eISSN - 2042-7158
pISSN - 0022-3573
DOI - 10.1211/jpp.61.02.0007
Subject(s) - sulfation , cytosol , recombinant dna , in vitro , piceatannol , biochemistry , chemistry , pharmacology , biology , enzyme , resveratrol , gene
Abstract Objectives The aim of this study was to investigate the concentration‐dependent sulfation of piceatannol, a dietary polyphenol present in grapes and wine and known for its promising anticancer and anti‐inflammatory activity. Methods Sulfation of piceatannol was investigated in human liver cytosol as well as using a panel of recombinant sulfotransferase isoforms. Furthermore, the chemical structures of novel sulfates were identified by liquid chromatography/mass spectrometry (LC/MS). Key findings In the presence of 3′‐phosphoadenosine‐5′‐phosphosulfate, three metabolites could be detected whose structures were identified by LC/MS/MS as piceatannol disulfate (M1) and two monosulfates (M2, M3). The kinetics of M1 formation exhibited a pattern of substrate inhibition with a K i of 21.8 ± 11.3 μM and a V max /K m of 7.63 ± 1.80 μl/mg protein per min. Formation of M2 and M3 showed sigmoidal kinetics with apparent K m and V max values of 27.1 ± 2.90 μM and 118.4 ± 4.38 pmol/mg protein per min, respectively, for M2; and 35.7 ± 2.70 μM and 81.8 ± 2.77 pmol/mg protein per min, respectively, for M3. Incubation in the presence of human recombinant sulfotransferases (SULTs) demonstrated that M1 was formed equally by SULT1A1*1 and SULT1B1 and to a lesser extent by SULT1A1*2. M2 was preferentially catalysed by SULT1A1*2, 1A3 and 1E1. The formation of M3, however, was mainly catalysed by SULT1A2*1 and SULT1A3. Conclusions Our results elucidate the importance of piceatannol sulfation in human liver, which must be taken into account in humans after dietary intake of piceatannol.