Skip to main content
SpringerLink
Log in

Expression of human liver 3, 4-catechol estrogens UDP-Glucuronosyltransferase cDNA in COS 1 cells

  • Research Articles
  • Published:
Archives of Pharmacal Research Aims and scope Submit manuscript

Abstract

The human cDNA clone UDPGTh2, encoding a liver UDP-glucuronosyltransferase (UDPGT), was isolated from a λgt 11 cDNA library by hybridization to mouse transferase cDNA clone, UDPGTm1. The two clones had 74% nucleotide sequence identities in the coding region UDPGTh2 encoded a 529 amino acid protein with an amino terminus membrane-insertion signal peptide and a carboxyl terminus membrane-spanning region. In order to establish substrate specificity, the clone was inserted into the pSVL vector (pUDPGTh2) and expressed in COS 1 cells. Sixty potential substrates were tested using cells transfected with pUDPGTh2. The order of relative substrate activity, was as follows: 4-hydroxyestrone > estriol >2-hydroxyestriol > 4-hydroxyestradiol > 6α-hydroxyestradiol > 5α-androstane-3α, 11β, 17β-triol=5β-androstane-3α, 11β, 17β-triol. There were only trace amounts of glucuronidation of 2-hydroxyestradiol and 2-hydroxyestrone, and in contrast to other cloned transferase, no gulcuronidation of either the primary estrogens and androgens (estrone, 17β-estradiol/testosterone, androsterone) or any of the exogenous substrates tested was detected. A lineweaver-Burk plot of the effect of 4-hydroxyestrone concentration on the velocity of glucuronidation showed an apparent Km of 13 μM. The unique specificity of this transferase might play an important role in regulating the level and activity of these potent and active estrogen metabolites.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References Cited

  • Bock, K. W., Lienblum, W. and Pfiel, H., Functional Heterogeneity of UDP-Glucuronosyltransferase activities in C57BL/6 and DBA/2 Mice.Biochem. Pharmacol., 31, 1273–1277 (1982).

    Article  PubMed  CAS  Google Scholar 

  • Church, G. and Gilbert, W. J., Genomic sequencing.Proc. Natl. Acad. Sci. U.S.A., 81, 1991–1995 (1984).

    Article  PubMed  CAS  Google Scholar 

  • Classen, E. and Doull A., Biotransformation of xenobiotics: Cassaret & Doull's toxicology, MacMillan P.C., New York, pp. 113–198, 1996.

    Google Scholar 

  • Dong, M. S., Owens, I. S. and Sheen, Y. Y., Cloning and expression of human liver UDP-glucuronosyl-transferase cDNA, UDPGTh2.Arch. Pharm. Res., 20, in press (1997).

  • Dutton, G. J.,Glucuronidation of drugs and other compounds. CRC Inc., Florida USA pp. 69–78, 1980.

    Google Scholar 

  • Fournel-Gigleux, S., Jackson, M. R., Wooster, R. and Burchell, B., Expression of a human liver cDNA encoding a UDP-glucuronosyltransferas catalysing the glucuronidation of hyodeoxycholic acid in cell culture.FEBS Lett., 243, 119–122 (1989).

    Article  PubMed  CAS  Google Scholar 

  • Harding, D., Fournel-Gigleux, S., Jackson, M. R. and Burchelle, B., Cloning and Substrate Specificity of a human phenol UDP-glucuronosyltransferase expressed in COS-7 cells.Proc. Natl. Acad. Sci. U.S.A., 85, 8381–8385 (1988).

    Article  PubMed  CAS  Google Scholar 

  • Horwitz, K. B. and McGuire, W. L., Estrogen control of progesterone receptor in human breast cancer.J. Biol. Chem., 253, 2223–2228 (1978b).

    PubMed  CAS  Google Scholar 

  • Horwitz, K. B. and McGuire, W. L., Antiestrogen mechanisms of action and effects in breast cancer: Breast cancer advances in research and treatment. Plenum Press, New York vol 2, pp. 155–204, 1978a.

    Google Scholar 

  • Jackson, M. R., McCarthy, L. R., Harding, D., Wilson, S., Coughtrie, M. W. H. and burchell, B., Cloning of a human liver microsomal UDP-glucurosyltransferase cDNA.Biochem. J., 242, 581–588 (1987).

    PubMed  CAS  Google Scholar 

  • Katzenellenbogen, B. S., Miller, M. A., Mullick, A. and Sheen, Y. Y., Antiestrogen action in breast cancer cells: Modulation of proliferation and protein synthesis and interaction with estrogen receptors and additional antiestrogen binding sites.Cancer Res. Treat., 5, 231–243 (1985).

    Article  CAS  Google Scholar 

  • Katzenellenbogen B. S., Norman, M. J., Eckert, R. L., Peltz, S. W. and Mangle, W. F., Bioactive, estrogen receptor interactions, and plasminogen activator-inducing activities of tamoxifen and hydroxytamoxifen in human breast cancer cells.Cancer Res., 44, 112–119 (1984).

    PubMed  CAS  Google Scholar 

  • Katzenellenbogen, B. S., Bhakoo, H. S., Fergson, E. R., Lan, N. C., Tatte, T., Tsai, T. L. and Katzenellenbogen, J. A., Estrogen and antiestrogen action in reproductive tissues and tumors.Recent Pro. Hormon Res., 35, 259–300 (1979).

    CAS  Google Scholar 

  • King, W. J. and Greene, G. J., Monoclonal antibodies localize estrogen receptor in nuclei of target cells.Nature (Lodon), 307, 745–747 (1984).

    Article  CAS  Google Scholar 

  • Kimura, T. and Owens I. S., Mouse UDP glucuronosyltransferase cDNA and complete amino acid sequence and regulation.Eur. J. Biochem., 168, 515–521 (1987).

    Article  PubMed  CAS  Google Scholar 

  • Kirkpatrick, R. B., Falany, C. N. and Tephly, T. R., Glucuronidation of Bile Acids by Rat Liver 3-OH Androgen UDP-Glucuronyltransferase.J. Biol. Chem., 259, 6176–6180 (1984).

    PubMed  CAS  Google Scholar 

  • Kornfeld, R. and Kornfeld, S., Assembly of asparagin-linked oligosaccharides.Ann. Rev. Biochem., 50, 555–583 (1981).

    Article  Google Scholar 

  • Mackenzie, P. I., Hjelmeland, L. H. and Owens, I. S., Purification and Immunochemical Characterization of a Low Form of UDP Glucuronosyltransferase from Mouse Liver.Arch. Biochem. Biophys., 231, 487–497 (1984).

    Article  PubMed  CAS  Google Scholar 

  • Mackenzie, P. I., Rat liver UDP-glucuronosyltransferase.J. Biol. Chem., 262, 9744–9749 (1986a).

    Google Scholar 

  • Mackenzie P. I., Rat liver UDP-glucuronosyltransferase.J. Biol. Chem., 161, 14112–14117 (1986b).

    Google Scholar 

  • Matern, H., Matern, S. and Gerok, W., Isolation and Characterization of rat Liver Microsomal UDP-Glucuronosyltransferase Activity toward Chenodeoxycholic Acid and Testosterone as a Single Form of Enzyme.J. Biol. Chem., 257, 7422–7429 (1982).

    PubMed  CAS  Google Scholar 

  • Miller, M. A. and Katzenellenbogen, B. S., Characterization and quantitation of antiestrogen binding sites in estrogen receptor-positive and-negative human breast cancer cell lines.Cancer Res., 43, 3094–3099 (1983).

    PubMed  CAS  Google Scholar 

  • Rorke, E. A. and katzenellenbogen B. S., Effects of estrogen and antiestrogens on estrogen receptor dynamics and the induction of progesterone receptor in MCF-7 human breast cancer cells.Cancer Res., 42, 139–144 (1982).

    Google Scholar 

  • Rose, T. K., Welsch, W. J., Sefton, B. M., Esch, F. S. and Ling N. C., Vesicular Stomatitis virus glycoprotein is anchored in the viral membrane by a hydrophobic domain near the COOH terminus.Proc. Natl. Acad. Sci. U.S.A., 77, 3884–3888 (1980).

    Article  PubMed  CAS  Google Scholar 

  • Sheen, Y. Y. and Katzenellenbogen B. S., Antiestrogen stimulation of the a 37,000 molecular weight secreted protein and estrogen stimulation of the 32,000 molecular weight secreted protein in MCF-7 human breast cancer cells.Endocrinol., 120, 1140–1151 (1987).

    Article  CAS  Google Scholar 

  • Sheen, Y. Y., Ruh, T. S., Mangel, W. F. and Katzenellenbogen B. S., An antiestrogenic potency and binding characteristics of the triphenylethylene H1285 in MCF-7 human breast cancer cells.Cancer Res., 45, 4192–4199 (1984).

    Google Scholar 

  • Sheen, Y. Y., Simpson, D. M. and Katzenellenbogen, B. S., An antiestrogen of the role of antiestrogen-binding sites in mediating the growth modulatory effects of antiestrogens: Studies using t-Butylphenoxyethyl diethylamine, a compound lacking affinity for the estrogen receptor.Endocrinol., 117, 561–564 (1985).

    CAS  Google Scholar 

  • Wishhart, G. J., Functional heterogenesity of UDP-glucuronosyltransferase as indicated by its differential development and inducibility by glucocorticoid.Biochem. J., 174, 485–489 (1978).

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Section of Drug Metabolism, Heritable Disease Branch, National Institute of Child and Human Development, National Institute of Health, USA

    Ida S. Owens

  2. College of Pharmacy, Ewha Womans University, #11-1, Daehyundong, Sudaemunku, 120-750, Seoul, Korea

    Mee Ryung Ahn & Yhun Yhong Sheen Ph.D.

Authors
  1. Mee Ryung Ahn
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ida S. Owens
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yhun Yhong Sheen Ph.D.
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Ahn, M.R., Owens, I.S. & Sheen, Y.Y. Expression of human liver 3, 4-catechol estrogens UDP-Glucuronosyltransferase cDNA in COS 1 cells. Arch. Pharm. Res. 20, 465–470 (1997). https://doi.org/10.1007/BF02973941

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02973941

Key words

Search

Navigation