title: Human UDP-glucuronosyltransferase 1A5: identification, expression, and activity.
abstract: The human UDP-glucuronosyltransferase (UGT) subfamily 1A includes nine genes. The expression of all the UGT1A isoforms, apart from UGT1A5, has been reported previously. We have now detected a low basal level of UGT1A5 expression in cultured human hepatocytes, and treatment with rifampicin or 3-methylcholanthrene increased the level of UGT1A5 mRNA. Low-level UGT1A5 expression was also found in HepG2 and Caco-2 cells as well as human liver. Furthermore, UGT1A5 expression has been detected in various segments of the intestine from human donors, revealing high interindividual variability in its level and distribution along the intestine. Full-length UGT1A5 cDNA was isolated from Caco-2 cells that had been transfected with the pregnane X receptor and treated with rifampicin. Recombinant UGT1A5, expressed in baculovirus-infected insect cells, exhibited very low rates of 4-methylumbelliferone and scopoletin glucuronidation, whereas 1-hydroxypyrene was a much better substrate for it. UGT1A5 did not glucuronidate 4-aminobiphenyl, a good substrate for the highly homologous enzymes UGT1A4 and UGT1A3. However, replacing the first 110 amino acids of UGT1A5, a region that may be involved in substrate binding, with the counterpart segment from UGT1A4 did not increase the 4-aminobiphenyl glucuronidation activity. Collectively, this work demonstrates for the first time that the human UGT1A5 is expressed in several tissues, is inducible, and is catalytically active.
authors.affiliation: Viikki DDTC, Faculty of Pharmacy, University of Helsinki, P.O. Box 56 (Viikinkaari 5E), 00014 University of Helsinki, Finland. moshe.finel@helsinki.fi
authors.forename: Moshe
authors.initials: M
authors.lastname: Finel
authors.forename: Xin
authors.initials: X
authors.lastname: Li
authors.forename: Dione
authors.initials: D
authors.lastname: Gardner-Stephen
authors.forename: Stacie
authors.initials: S
authors.lastname: Bratton
authors.forename: Peter I
authors.initials: PI
authors.lastname: Mackenzie
authors.forename: Anna
authors.initials: A
authors.lastname: Radominska-Pandya
mesh_terms: D000328:Adult; D000368:Aged; D000595:Amino Acid Sequence; D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D018938:Caco-2 Cells; D006528:Carcinoma, Hepatocellular / Q000201:enzymology* / Q000473:pathology; D002384:Catalysis; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D018076:DNA, Complementary; D015870:Gene Expression; D014453:Glucuronosyltransferase / Q000032:analysis / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D022781:Hepatocytes / Q000187:drug effects / Q000201:enzymology*; D006801:Humans; D007313:Insecta / Q000166:cytology / Q000821:virology; D007413:Intestinal Mucosa / Q000201:enzymology; D007527:Isoenzymes / Q000032:analysis / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D008113:Liver Neoplasms / Q000201:enzymology* / Q000473:pathology; D008297:Male; D008748:Methylcholanthrene / Q000494:pharmacology; D008875:Middle Aged; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D012293:Rifampin / Q000494:pharmacology; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity
chemical_list: D018076:DNA, Complementary; D007527:Isoenzymes; D012333:RNA, Messenger; D011994:Recombinant Proteins; D008748:Methylcholanthrene; D014453:Glucuronosyltransferase; C507232:UGT1A5 protein, human; D012293:Rifampin