Cigarette smoke toxicants as substrates and inhibitors for human cytosolic SULTs.
The current study was designed to examine the role of sulfation in the metabolism of cigarette smoke toxicants and clarify whether these toxicants, by serving as substrates for the cytosolic sulfotransferases (SULTs), may interfere with the sulfation of key endogenous compounds. By metabolic labeling, [(35)S]sulfated species were found to be generated and released into the media of HepG2 human hepatoma cells and primary human lung endothelial cells labeled with [(35)S]sulfate in the presence of cigarette smoke extract (CSE). Concomitantly, several [(35)S]sulfated metabolites observed in the medium in the absence of CSE either decreased or disappeared. Eleven previously prepared human cytosolic SULTs were tested for sulfating activity with CSE and known cigarette smoke toxicants as substrates. Activity data revealed SULT1A1, SULT1A2, SULT1A3, and SULT1C#2 as major enzymes responsible for their sulfation. To examine their inhibitory effects on the sulfation of 17beta-estradiol by SULT1A1, enzymatic assays were performed in the presence of three representative toxicant compounds, namely N-hydroxy-4-aminobiphenyl (N-OH-4-ABP), 4-aminobiphenyl (4-ABP) and 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP). IC(50) values determined for the sulfation of 17beta-estradiol by SULT1A1 were 11.8 microM, 28.2 microM, and 500 microM, respectively, for N-OH-4-ABP, 4-ABP and PhIP. Kinetic analyses indicated that the mechanism underlying the inhibition of 17beta-estradiol sulfation by these cigarette smoke toxicants is of a mixed competitive-noncompetitive type. Metabolic labeling experiments clearly showed inhibition of the production of [(35)S]sulfated 17beta-estradiol by N-OH-4-ABP in a concentration-dependent manner in HepG2 cells. Taken together, these results suggest that sulfation plays a significant role in the metabolism of cigarette smoke compounds. By serving as substrates for SULTs, cigarette smoke toxicants may interfere with the metabolism of 17beta-estradiol and other endogenous compounds.
Yasuda
Shin
S
Biomedical Research Center, The University of Texas Health Center, 11937 US Highway 271, Tyler, TX 75708, USA.
Idell
Steven
S
Fu
Jian
J
Carter
Glendora
G
Snow
Rhodora
R
Liu
Ming-Cheh
MC
D000611:Aminobiphenyl Compounds / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D015239:Arylsulfotransferase / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D002855:Chromatography, Thin Layer / Q000379:methods; D003600:Cytosol / Q000201:enzymology*; D004305:Dose-Response Relationship, Drug; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D004958:Estradiol / Q000378:metabolism; D004965:Estrogen Antagonists / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D006801:Humans; D007093:Imidazoles / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D012906:Smoke / Q000032:analysis*; D013379:Substrate Specificity; D013431:Sulfates / Q000737:chemistry / Q000378:metabolism; D013462:Sulfur Radioisotopes; D014026:Nicotiana / Q000737:chemistry*
D000611:Aminobiphenyl Compounds; D004791:Enzyme Inhibitors; D004965:Estrogen Antagonists; D007093:Imidazoles; D012906:Smoke; D013431:Sulfates; D013462:Sulfur Radioisotopes; C006757:4-biphenylamine; D004958:Estradiol; C014593:N-hydroxy-4-aminobiphenyl; C049584:2-amino-1-methyl-6-phenylimidazo(4,5-b)pyridine; D015239:Arylsulfotransferase; C117544:SULT1A1 protein, human