Archives of Pharmacal Research

, Volume 20, Issue 5, pp 459–464 | Cite as

Cloning and expression of human liver UDP-glucuronosyltransferase cDNA, UDPGTh2

  • Misook Dong
  • Ida S. Owens
  • Yhun Yhong Sheen
Research Articles


The human liver cDNA clone UDPGTh2, encoding a liver UDP-glucuronosyltransferase (UDPGT) was isolated from a λ 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 [35S]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.

Key words

UDP-Glucuronosyltransferase cDNA UDPGTh2 Tunicamycin 


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Copyright information

© The Pharmaceutical Society of Korea 1997

Authors and Affiliations

  • Misook Dong
    • 2
  • Ida S. Owens
    • 1
  • Yhun Yhong Sheen
    • 2
  1. 1.Section of Drug Metabolism, Heritable Disease Branch, National Institute of Child and Human DevelopmentNational Institute of HealthUSA
  2. 2.College of PharmacyEwha Womans UniversitySeoulKorea

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