Use of receptor antibodies to demonstrate membrane glucocorticoid receptor in cells from human leukemic patients

Bahiru Gametchu, Cheryl S. Watson, Shenhong Wu

    Research output: Contribution to journalArticle

    101 Citations (Scopus)

    Abstract

    Anti-peptide antibody to the human glucocorticoid receptor (GR) was produced and used to demonstrate that a subset of the GR population resides in the plasma membrane of human leukemic cells. Characterization of the antibody with intracellular GR (iGR) showed its ability to shift [3H]triamcinolone acetonidelabeled GR (4S protein) from two human leukemic cell lines to a higher density in sucrose gradients; Western and autoradiographic analysis of affinity-labeled ([3H]dexamethasone 21-mesylate) receptor revealed an immunoreactive and competitively labeled band of 94 kDa. CCRF-CEM cell membrane GR (mGR) resolved as a > 7S protein on density gradients and immunoselected cell surface protein labeled by whole cell biotinylation or affinity-labeling with [3H]dexamethasone 21-mesylate was -145 kDa, demonstrating that mGR was larger in size than iGR, as has been shown previously for the mGR of mouse lymphoma cells. Analysis of mGR in lymphocytes of leukemic patients and the CCRF-CEM cell line indicated differences in levels of expression as shown by FACS and immunocytochemical analyses. We are currently using this system to study the correlation between the quantity of membrane-resident GRs and the glucocorticoid-induced lytic response, a relationship previously shown in the murine (S-49 cell) system.

    Original languageEnglish (US)
    Pages (from-to)1283-1292
    Number of pages10
    JournalFASEB Journal
    Volume7
    Issue number13
    StatePublished - Oct 1993

    Fingerprint

    Glucocorticoid Receptors
    Membranes
    receptors
    antibodies
    Antibodies
    Cell membranes
    dexamethasone
    cells
    Cells
    triamcinolone
    Cell Membrane
    biotinylation
    Biotinylation
    line differences
    Triamcinolone
    Cell Line
    human cell lines
    Lymphocytes
    mice
    surface proteins

    Keywords

    • Leukemia
    • Lymphocytolysis
    • Membrane glucocorticoid receptor
    • Steroid

    ASJC Scopus subject areas

    • Agricultural and Biological Sciences (miscellaneous)
    • Biochemistry, Genetics and Molecular Biology(all)
    • Biochemistry
    • Cell Biology

    Cite this

    Use of receptor antibodies to demonstrate membrane glucocorticoid receptor in cells from human leukemic patients. / Gametchu, Bahiru; Watson, Cheryl S.; Wu, Shenhong.

    In: FASEB Journal, Vol. 7, No. 13, 10.1993, p. 1283-1292.

    Research output: Contribution to journalArticle

    Gametchu, Bahiru ; Watson, Cheryl S. ; Wu, Shenhong. / Use of receptor antibodies to demonstrate membrane glucocorticoid receptor in cells from human leukemic patients. In: FASEB Journal. 1993 ; Vol. 7, No. 13. pp. 1283-1292.
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