title: Gastrointestinally distributed UDP-glucuronosyltransferase 1A10, which metabolizes estrogens and nonsteroidal anti-inflammatory drugs, depends upon phosphorylation.
abstract: Among gastrointestinal distributed isozymes encoded at the UGT1 locus, UDP-glucuronosyltransferase 1A10 (UGT1A10) metabolizes a number of important chemicals. Similar to broad conversion of phytoestrogens (Basu, N. K., Ciotti, M., Hwang, M. S., Kole, L., Mitra, P. S., Cho, J. W., and Owens, I. S. (2004) J. Biol. Chem. 279, 1429-1441), UGT1A10 metabolized estrogens and their derivatives, whereas UGT1A1, -1A3, -1A7, and -1A8 differentially exhibited reduced activity toward the same. UGT1A10 compared with UGT1A7, -1A8, and -1A3 generally exhibited high activity toward acidic nonsteroidal anti-inflammatory drugs and natural benzaldehyde derivatives, while UGT1A3 metabolized most efficiently aromatic transcinnamic acids known to be generated from flavonoid glycosides by microflora in the lower gastrointestinal tract. Finally UGT1A10, -1A7, -1A8, and -1A3 converted plant-based salicylic acids; methylsalicylic acid was transformed at high levels, and acetylsalicylic (aspirin) and salicylic acid were transformed at moderate to low levels. Atypically UGT1A10 transformed estrogens between pH 6 and 8 but acidic structures preferentially at pH 6.4. Furthermore evidence indicates UGT1A10 expressed in COS-1 cells depends upon phosphorylation; UGT1A10 versus its single, double, and triple mutants at three predicted protein kinase C phosphorylation sites incorporated [(33)P]-orthophosphate and showed a progressive decrease with no detectable label or activity for the triple T73A/T202A/S432G-1A10 mutant. Single and double mutants revealed either null/full activity or null/additive activity, respectively. Additionally UGT1A10-expressing cultures glucuronidated 17beta-[(14)C]estradiol, whereas cultures containing null mutants at protein kinase C sites showed no estrogen conversion. Importantly UGT1A10 in cells supported 10-fold higher glucuronidation of 17beta-estradiol than UGT1A1. In summary, our results suggest gastrointestinally distributed UGT1A10 is important for detoxifying estrogens/phytoestrogens and aromatic acids with complementary activity by UGT1A7, -1A8, -1A3, and/or -1A1 evidently dependent upon phosphorylation.
authors.affiliation: Heritable Disorders Branch, National Institute of Child Health & Human Development, National Institutes of Health, Room 9S-21, Building 10, Bethesda, MD 20892, USA.
authors.forename: Nikhil K
authors.initials: NK
authors.lastname: Basu
authors.forename: Shigeki
authors.initials: S
authors.lastname: Kubota
authors.forename: Meselhy R
authors.initials: MR
authors.lastname: Meselhy
authors.forename: Marco
authors.initials: M
authors.lastname: Ciotti
authors.forename: Bhabadeb
authors.initials: B
authors.lastname: Chowdhury
authors.forename: Masao
authors.initials: M
authors.lastname: Hartori
authors.forename: Ida S
authors.initials: IS
authors.lastname: Owens
mesh_terms: D000818:Animals; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000494:pharmacology*; D001665:Binding Sites; D015153:Blotting, Western; D019556:COS Cells; D002460:Cell Line; D002934:Cinnamates / Q000737:chemistry; D018076:DNA, Complementary / Q000378:metabolism; D004064:Digestive System / Q000201:enzymology*; D004305:Dose-Response Relationship, Drug; D004958:Estradiol / Q000378:metabolism; D004967:Estrogens / Q000378:metabolism*; D014453:Glucuronosyltransferase / Q000378:metabolism*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008861:Microsomes / Q000378:metabolism; D008956:Models, Chemical; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D010766:Phosphorylation; D011493:Protein Kinase C / Q000378:metabolism; D012459:Salicylates / Q000378:metabolism; D014018:Tissue Distribution; D014162:Transfection
chemical_list: D000894:Anti-Inflammatory Agents, Non-Steroidal; D002934:Cinnamates; D018076:DNA, Complementary; D004967:Estrogens; D012459:Salicylates; C029010:cinnamic acid; D004958:Estradiol; C122075:UDP-glucuronosyltransferase, UGT1A3; C476725:bilirubin uridine-diphosphoglucuronosyl transferase 1A10; D014453:Glucuronosyltransferase; C107611:UDP-glucuronosyltransferase, UGT1A7; C454320:UDP-glucuronosyltransferase, UGT1A8; D011493:Protein Kinase C