Proteins pp 61-74 | Cite as

High-Performance Liquid Chromatography as a Means of Characterizing Isoforms of Steroid Hormone Receptor Proteins

  • J. L. Wittliff
  • N. A. Shahabi
  • S. M. Hyder
  • L. A. van der Walt
  • L. Myatt
  • D. M. Boyle
  • Y.-J. He

Abstract

Steroid hormone receptors remain one of the most elusive, labile proteins under study in the field of molecular endocrinology. To our knowledge, no one has provided conclusive evidence of the “native state” of either estrogen or progestin receptors in breast and endometrial carcinomas. Most investigators have utilized sucrose gradient centrifugation, conventional column chromatography and/or isoelectric focusing to characterize these proteins in impure preparations1,2. Using these techniques, several receptor species have been identified.

Keywords

HPLC Glycerol Acetonitrile Sedimentation Tamoxifen 

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References

  1. 1.
    J.L. Wittliff, Steroid Binding Proteins in Normal and Neoplastic Mammary Cells, in:“Methods in Cancer Research,” H. Busch led.), Vol. XI:293–354, New York:Academic Press, 1975.Google Scholar
  2. 2.
    J.L. Wittliff and R.D. Wiehle, Analytical methods for steroid hormone receptors and their quality assurance, in:“Hormonally Sensitive Tumors,” V.P. Hollander, ed., Academic Press, Inc. (in press).Google Scholar
  3. 3.
    J.L. Wittliff, P.W. Feldhoff, A. Fuchs, and R.D. Wiehle, Polymorphism of estrogen receptors in human breast cancer Polymorphism of estrogen receptors in human breast cancer, in:“Physiology of Endocrine Disease and Mechanisms of Hormone Action,” R.J. Soto, A.F. Denicola, and J.A. Blaquier, eds., Alan R. Liss, Inc., New York (1981).Google Scholar
  4. 4.
    J.L. Wittliff, Separation and characterization of isoforms of steroid hormone receptors using high-performance liquid chromatography, in:“Molecular Mechanisms of Steroid Hormone Action,” V.K. Moudgil, ed., Walter de Gruyter and Co., Berlin, Germany, (1985).Google Scholar
  5. 5.
    R.D. Wiehle, G.E. Hofmann, A. Fuchs, and J.L. Wittliff, High-performance size exclusion chromatography as a rapid methods for the separation of steroid hormone receptor. J. Chromatogr. 307:39 (1984).Google Scholar
  6. 6.
    N.A. Shahabi, T.W. Hutchens, J.L. Wittliff, M.E. Kirk, S.D. Halmo, and J.A. Nisker, Physicochemical characterization of estrogen receptors from a rabbit endometrial carcinoma model. Prog. Cancer Res. & Therap. 31:63 (1984).Google Scholar
  7. 7.
    R.D. Wiehle and J.L. Wittliff, Isoforms of estrogen receptors by high-performance ion-exchange chromatography. J. Chromatogr. 297:313 (1984).PubMedCrossRefGoogle Scholar
  8. 8.
    D.M. Boyle, R.D. Wiehle, N.A. Shahabi, and J.L. Wittliff, A rapid high-resolution procedure for assessment of estrogen receptor heterogeneity in clinical samples. J. Chromatogr. 327:369 (1985).PubMedCrossRefGoogle Scholar
  9. 9.
    L.A. van der Walt and J.L. Wittliff, High resolution separation of molybdate-stabilized progestin receptors using high performance liquid chromatography. J. Chromatogr. (in press).Google Scholar
  10. 10.
    L.A. van der Walt and J.L. Wittliff Assessment of progestin receptor polymorphism by various synthetic ligands using HPLC, J. Steroid Biochem. (in press).Google Scholar
  11. 11.
    L. Myatt and J.L. Wittliff, Characterization of non activated and activated estrogen and antiestrogen receptor complex by high performance ion-exchange chromatography. 67th Annual Meeting, The Endocrine Society, 1156 (1985).Google Scholar
  12. 12.
    N.A. Shahabi, R.D. Wiehle, S.M. Hyder, and J.L. Wittliff, Analysis of steroid hormone receptors by multi-dimensional approach utilizing high-performance liquid chromatography (HPLC) methodologies, IV Intl. Symp. HPLC of Proteins, Peptides, and Polynecleotides Abs. (1984).Google Scholar
  13. 13.
    T.W. Hutchens, R.D. Wiehle, N.A. Shahabi, and J.L. Wittliff, Rapid analysis of estrogen receptor heterogeneity by chromatofocusing with high-performance liquid chromatography, J. Chromatogr. 266:115 (1983).CrossRefGoogle Scholar
  14. 14.
    S.M. Hyder, R.D. Wiehle, D.W. Brandt, and J.L. Wittliff, High-performance hydrophobic-interaction chromatography of steroid hormone receptors, J. Chromatogr. 32:237 (1985).CrossRefGoogle Scholar
  15. 15.
    M. Lonsdorfer, N.C. Clements, Jr., and J.L. Wittliff, Performance Liquid Chromatography in the Elevation of the Synthesis and Binding of Fluorescein-Linked Steroids to Estrogen Receptors., J. Chromatogr. 266:129, 1983.PubMedCrossRefGoogle Scholar
  16. 16.
    L.S. Dure IV, W.T. Schrader, and B.W. O’Malley, Covalent attachment of a progestational Steroid to chick oviduct progesterone receptor by photoaffinity labelling, Nature 283:784, 1980.Google Scholar
  17. 17.
    G.L. Greene, C. Nolan, J.P. Englar and E.V. Jensen, Monoclonal Antibodies to human estrogen receptor, Proc. Natl. Acad. Sci, USA 77:5115, 1980.PubMedCrossRefGoogle Scholar
  18. 18.
    W.J. Waddell, A Simple ultraviolet spectrophotmetric method for determining of proteins. J. Lab. Clin. Med., 48:311, 1956.PubMedGoogle Scholar
  19. 19.
    M.M. Bradford, A rapid and sensitve method for the quantitation of microgram quantities of protein utilizing the principle of protein dye binding. Anal. Biochem. 72:248, 1976.PubMedCrossRefGoogle Scholar
  20. 20.
    N. Sato, S.M. Hyder, L. Chang, A. Thais, and J.L. Wittliff Interaction of Estrogen Receptor Isoforms with Immobilized Monoclonal Antibodies, V Intl. Symp. HPLC of Proteins, Peptides, and Polynecleotides Abs. (1985).Google Scholar
  21. 21.
    S.M. Hyder, R.D. Wiehle, and J.L. Wittliff, Alterations in the estrogen receptor isoforms in the breast and uterus of the rat during differentiation, Fed. Proc. 44:1474 (1985).Google Scholar

Copyright information

© Plenum Press, New York 1987

Authors and Affiliations

  • J. L. Wittliff
    • 1
    • 2
  • N. A. Shahabi
    • 1
    • 2
  • S. M. Hyder
    • 1
    • 2
  • L. A. van der Walt
    • 1
    • 2
  • L. Myatt
    • 1
    • 2
  • D. M. Boyle
    • 1
    • 2
  • Y.-J. He
    • 1
    • 2
  1. 1.Hormone Receptor Laboratory, Department of BiochemistryJames Graham Brown Cancer CenterUSA
  2. 2.University of Louisville School of MedicineLouisvilleUSA

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