Heterogeneity of AtT-20 Cell Glucocorticoid Binding Sites: Evidence for a Membrane Receptor

  • R. W. Harrison
  • K. Balasubramanian
  • J. Yeakley
  • M. Fant
  • F. Svec
  • S. Fairfield
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 117)


In previous studies we have found that intact AtT-20 cells contained two glucocorticoid binding sites with distinctly different affinities and specificity. In this paper, the nature of these sites was investigated by studying glucocorticoid binding to cytosol and to plasma membranes isolated from AtT-20 mouse pituitary tumor cells. Plasma membrane vesicles were isolated from AtT-20 cells and found to take up α-aminoisobutyric acid, indicating that they were properly oriented and functionally intact. Corticosterone bound to these vesicles in a time- and temperature-dependent manner. The binding exhibited a glucocorticoid preference since non-glucocorticoids such as progesterone, testosterone or estradiol were unable to inhibit binding. In addition, binding specificity differed from that of the cytoplasmic receptor since the synthetic glucocorticoids were also ineffective competitors. The major inhibitors of binding were corticosterone > 11-dehydrocorticosterone > 11-ketoprogesterone > cortisol. In other complementary studies, AtT-20 cell cytosol was tested to determine whether heterogenous soluble sites exhibiting a preference for the natural vs the synthetic steroid could also be identified. We found that binding sites for both steroid classes were approximately similar in number, specificity and behavior on ion-exchange chromatography.

We conclude that, in addition to a classical soluble cytoplasmic glucocorticoid receptor, AtT-20 cells contain plasma membrane glucocorticoid binding sites. The affinity and specificity of these sites for the natural ligand, corticosterone, suggest that they play an important role in the subcellular mechanism of glucocorticoid action.


Glucocorticoid Receptor Triamcinolone Acetonide Synthetic Glucocorticoid Relative Binding Affinity Cytosol Receptor 
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Copyright information

© Springer Science+Business Media New York 1979

Authors and Affiliations

  • R. W. Harrison
    • 1
  • K. Balasubramanian
    • 1
  • J. Yeakley
    • 1
  • M. Fant
    • 1
  • F. Svec
    • 1
  • S. Fairfield
    • 1
  1. 1.Department of Medicine and BiochemistryVanderbilt University School of MedicineNashvilleUSA

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