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The Pharmaceutical Society of Korea (대한약학회)
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Cloning and Expression of Human Liver UDP-Glucuronosyltransferase cDNA, UDPGTh2

  • Dong, Misook (College of Pharmacy, Ewha Womans University) ;
  • Owens, Ida-S. (Section Of Drug Metabolism Heritable Disease Branch, National Institute of Child and Human Development, National Institute of Health) ;
  • Sheen, Yhun-Yhong (College of Pharmacy, Ewha Womans University)
  • Published : 1997.10.01
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Abstract

The human liver cDNA clone UDPGTh2, encoding a liver UDP-glucuronosyltransferase (UDPGT) was isolated from a .gamma. gt 11 cDNA library by hybridization to mouse transferase cDNA clone, UDPGTm1. UDPGTh2 encoded a 529 amino acid protein with an amino terminus membrane-insertion signal peptide and a carboxyl terminus membrane-spanning region. There were three potential asparagine-linked glycosylation sites at residues 67, 68, and 315. In order to obtain UDPGTh2 protein encoded from cloned human liver UDP-glucuronosyltransferase cDNA, the clone was inserted into the pSVL vector (pUDPGTh2) and expressed in COS 1 cells. The presence of a transferase with Mr-52,000 in transfected cells cultured in the presence of $[^{35}S]$ methionine was shown by immunocomplexed products with goat antimouse transferase IgG and protein A-Sepharose and analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography. The expressed UDPGT was a glycoprotein as indicated by electrophoretic mobility shift in Mr-3,000-4,000 when expressed in the presence of tunicamycin. The extent of glycosylation was difficult to assess, although one could assume that glycosyl structures incorporated at the level of endoplasmic reticulum were always the core oligosaccharides. Thus, it is likely that at least two moieties inserted can account for the shift of Mr-3,000-4,000. This study demonstrates the cDNA and deduced amino acid sequence of human liver UDP-glucuronosyltransferase cDNA, UDPGTh2.

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References

  1. Biochem. Pharm. v.27 UDP-glucuronosyltransferase in perfused rat liver and in microsomes Bock,W.K.;Clausbruch U.C.V.;Ottenwalder H.
  2. Biochem. Pharmacol v.31 Functional Heterogeneity of UDP-Glucuronosyltransferase activities in C57BL/6 and DBA/2 Mice Bock,K.W.;Lilienblum,W.;Pfiel,H.
  3. Biochem. J. v.223 Bilirubin mono- and diglucuronide formation by purified rat liver microsomal bilirubin UDP-glucuronosylransterase Burchell B.;Blakckaert,N.
  4. Proc. Natl. Acad. Sci. U.S.A. v.81 Genomic sequencing Church,G.;Gilbert,W.J.
  5. Glucuronidation of drugs and other compounds Dutton,G.J.
  6. FEBS Lett. v.243 Expression of a human liver cDNA encoding a UDP-glucuronosyltransferas catalysing the glucuronidation of hyodeoxycholic acid in cell culture Fournel-Gigleux,S.;Jackson,M.R.;Wooster,R.;Burchell,B.
  7. Proc. Natl. Acad. Sci. U.S.A. v.85 Cloning and Substrate Specificity of a human phenol UDP-glucuronosyltransferase expressed in COS-7 cells Harding,D.;Fournel-Gigleux,S.;Jackson,M.R.;Burchelle,B.
  8. Ann. Rev. Bichem. v.50 Assembly of asparaginlinked oligosaccharides Kornfeld.R.;Kornfeld.S.
  9. Biohem. J. v.242 Cloning of a human liver microsomal UDP-glucuronosyltransferase cDNA Jankson,M.R.;McCarthy,L.R.;Harding,D.;Wilson,S.;Coughtrie,M.W.H.;burchell,B.
  10. Eur. J. Biochem v.168 Mouse UDP glucuronosyltransferase cDNA and complete amino acid sequence and regulation Kimura,T.;Owens I.S.
  11. J. Biol. Chem. v.259 Glucuronidation of Bile Acids by Rat Liver 3-OH Androgen UDP-Glucuronosyltransferase Kirkpatrick,R.B.;Falany,C.N.;Tephly,T.R.
  12. Arch. Biochem. Biophys v.231 Purification and Immunochemical Characterization of a Low Form of UDP Glucuronosyltransferase from Mouse Liver Mackenzie,P.I.;Hjelmeland,L.H.;Owens,I.S.
  13. J. Biol. Chem. v.257 Isolation and Characterization of rat Liver Microsomal UDP-Glucuronosyltransferase Activity toward Chenodeoxycholic Acid and Testosterone as a Single Form of Enzyme Matern,H.;Matern,S.;Gerok,W.
  14. Proc. Natl. Acad. Sci. U.S.A. v.77 Vesicular Stomatitis virus glycoprotein is anchored in the viral membrane by a hydrophobic domain near the COOH terminus Rose,T.K.;Welsch,W.J.;Sefton,B.M.;Esch,F.S.;Ling,N.C.
  15. Biochem. J. v.174 Functional heterogenesity of UDP-glucuronosyltransferase as indicated by its differential development and inducibility by glucocorticoid Wishhart,G.J.