title: Stabilization of glucocorticoid-receptor complexes in rat thymus cytosol by a factor from WEHI-7 cells.
abstract: Using a variety of physico-chemical techniques we have recently characterized three distinct forms of glucocorticoid-receptor complexes present in the cytosol from rat thymus cells incubated with glucocorticoid; the relative proportions of these complexes are dependent on the conditions to which the cells or cytosols are exposed. Two of these complexes correspond to the well established nonactivated and activated receptor forms, while the third has properties consistent with mero-receptor. Based on their differential affinities for DNA- and DEAE-cellulose we have developed a rapid mini-column chromatographic procedure for separating these three forms and have used it to examine the stability of complexes in cytosol preparations. We have found that activated glucocorticoid-receptor complexes from rat thymus cells are relatively unstable under cell-free conditions in that they undergo time-dependent losses in DNA binding and are converted to mero-receptor. In contrast, cytosolic glucocorticoid-receptor complexes prepared from WEHI-7 mouse thymoma cells are remarkably stable under similar conditions. Mixing experiments with equal portions of rat thymus and WEHI-7 cytosol revealed that the difference between the two tissues cannot be accounted for merely by differences in amounts of proteolytic enzymes, since addition of rat thymus cytosol to WEHI-7 cytosol containing activated glucocorticoid-receptor complexes does not result in their conversion to mero-receptor. However, the WEHI-7 cytosol affords considerable protection to activated glucocorticoid-receptor complexes in thymus cytosol. The stabilizing factor from WEHI-7 cytosol is heat stable (survives 100 degrees C for 30 min), insensitive to pH over a wide range (4.0-10.0), and appears to be macromolecular. It does not inhibit activation, and thus appears distinct from the previously described endogenous glucocorticoid receptor stabilizing factor responsible for stabilization of thymocyte receptor binding capacity (Leach et al., J. Biol. Chem. 257: 381-388, 1982). We propose that the factor is an endogenous inhibitor of the protease(s) responsible for mero-receptor formation.
mesh_terms: D000818:Animals; D002460:Cell Line; D003600:Cytosol / Q000378:metabolism; D006801:Humans; D051379:Mice; D051381:Rats; D011965:Receptors, Glucocorticoid / Q000187:drug effects / Q000378:metabolism*; D011987:Receptors, Steroid / Q000378:metabolism*; D013696:Temperature; D013945:Thymoma / Q000032:analysis*; D013950:Thymus Gland / Q000378:metabolism*; D013953:Thymus Neoplasms / Q000032:analysis*; D014020:Tissue Extracts / Q000494:pharmacology*
chemical_list: D011965:Receptors, Glucocorticoid; D011987:Receptors, Steroid; D014020:Tissue Extracts