Sulfation of resveratrol in human liver: Evidence of a major role for the sulfotransferases SULT1A1 and SULT1E1

Abstract

Sulfation of resveratrol, a polyphenolic compound present in grapes and wine with anticancer and cardioprotective activities, was studied in human liver cytosol. In the presence of 3'-phosphoadenosine-5'-phosphosulfate, three metabolites (M1-3) whose structures were identified by mass spectrometry and NMR as trans-resveratrol-3-O-sulfate, trans-resveratrol-4'-O-sulfate, and trans-resveratrol-3-O-4'-O-disulfate, respectively. The kinetics of M1 formation in human liver cytosol exhibited an pattern of substrate inhibition with a K(i) of 21.3 +/- 8.73 microM and a V(max)/K(m) of 1.63 +/- 0.41 microLmin(-1)mg(-1) protein. Formation of M2 and M3 showed sigmoidal kinetics with about 56-fold higher V(max)/K(m) values for M3 than for M2 (2.23 +/- 0.14 and 0.04 +/- 0.01 microLmin(-1)mg(-1)). Incubation in the presence of human recombinant sulfotransferases (SULTs) demonstrated that M1 is almost exclusively catalysed by SULT1A1 and only to a minor extent by SULT 1A2, 1A3 and 1E1, whereas M2 is selectively formed by SULT1A2. M3 is mainly catalysed by SULT1A2 and 1A3. In conclusion, the results elucidate the enzymatic pathways of resveratrol in human liver, which must be considered in humans following oral uptake of dietary resveratrol.

para-Nitrophenyl Sulfate Activation of Human Sulfotransferase 1A1 Is Consistent with Intercepting the E·PAP Complex and Reformation of E·PAPS

“…However, the intercepts of the lines at these concentrations do exhibit activation by pNPS, and this reflects the second half of Reaction 3 i.e. the steps represented by rate constants k 11 to k 15 . Under these conditions, it is unlikely that release of PAP (Q) from E⅐PAP contributes to the overall reaction.…”

“…Substrate inhibition by the hydroxyl substrate (sulfate acceptor) is a common feature of most cytosolic SULTs (14,15). Inhibition of SULT1A1 has been observed by the substrate, naphthol.…”

“…V ϭ k 13 k 15 k 13 ϩ k 14 ϩ k 15 (Eq. 6) The activation by pNPS thus results from bypassing the release of PAP from E⅐PAP by direct sulfuryl transfer from pNPS to PAP on enzyme.…”

“…SULT1A1 is also widely distributed in the human body. On the basis of isotope exchange at equilibrium, we showed that the kinetic mechanism for human SULT1A1 is steady-state-ordered with PAPS binding to the protein first, and PAP released last (13).Substrate inhibition by the hydroxyl substrate (sulfate acceptor) is a common feature of most cytosolic SULTs (14,15). Inhibition of SULT1A1 has been observed by the substrate, naphthol.…”

Pulmonary Metabolism of Resveratrol: In Vitro and In Vivo Evidence

“…It also has lung cancer chemopreventive activity by altering the expression of genes involved in the phase 1 metabolism of polycyclic hydrocarbons (Mollerup et al, 2001). Although the role of the gut and liver is well known in the metabolism of RES (Kuhnle et al, 2000;Miksits et al, 2005;Brill et al, 2006;Iwuchukwu and Nagar, 2008;van de Wetering et al, 2009), the role of lungs has not been evaluated in the metabolism of RES. RES is known to be extensively metabolized into its two major metabolites [i.e., trans-resveratrol-3-sulfate (R3S) and trans-resveratrol-3-glucuronide (R3G)] (Fig.…”