Subunit composition of the untransformed glucocorticoid receptor in the cytosol and in the cell.
We have used bifunctional reagents to examine the subunit composition of the non-DNA-binding form of the rat and human glucocorticoid receptor. Treatment of intact cells and cell extracts with a reversible cross-linker, followed by electrophoretic analysis of immunoadsorbed receptor revealed that three proteins of apparent approximate molecular masses, 90, 53 and 14 kDa are associated with the receptor. The first of these was identified immunochemically as a 90-kDa heat-shock protein (hsp90). The complex isolated from HeLa cells contained 2.2 mol hsp90/mol steroid-binding subunit. Cross-linking of the receptor complex in the cytosol completely prevented salt-induced dissociation of the subunits. The cross-linked receptor was electrophoretically resolved into two oligomeric complexes of apparent molecular mass 288 kDa and 347 kDa, reflecting the association of the 53-kDa protein with a fraction of the receptor. Since no higher oligomeric complexes could be generated by cross-linking cell extracts under different conditions, we conclude that most of the untransformed cytosolic receptor is devoid of additional components.
Alexis
M N
MN
Institute of Biological Research and Biotechnology, National Hellenic Research Foundation, Athens, Greece.
Mavridou
I
I
Mitsiou
D J
DJ
D000818:Animals; D006528:Carcinoma, Hepatocellular; D002499:Centrifugation, Density Gradient; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D002845:Chromatography; D003432:Cross-Linking Reagents; D003600:Cytosol / Q000737:chemistry; D003907:Dexamethasone / Q000031:analogs & derivatives / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D006367:HeLa Cells; D006801:Humans; D015151:Immunoblotting; D008099:Liver / Q000737:chemistry; D008113:Liver Neoplasms; D046911:Macromolecular Substances; D008297:Male; D008970:Molecular Weight; D051381:Rats; D011919:Rats, Inbred Strains; D011965:Receptors, Glucocorticoid / Q000737:chemistry* / Q000378:metabolism; D013388:Succinimides; D014407:Tumor Cells, Cultured
D003432:Cross-Linking Reagents; D046911:Macromolecular Substances; D011965:Receptors, Glucocorticoid; D013388:Succinimides; C028722:ethylene glycolylbis(succinimidyl succinate); D003907:Dexamethasone; C011240:dithiobis(succinimidylpropionate); C031710:dexamethasone 21-methanesulfonate