Study of the heteromeric structure of the untransformed glucocorticoid receptor using chemical cross-linking and monoclonal antibodies against the 90K heat-shock protein.
The untransformed rat glucocorticoid receptor is assumed to be a hetero-oligomeric complex, containing a non-steroid binding component, the 90K heat-shock protein (HSP 90). Direct measurement of its molecular weight by chemical cross-linking provides new evidence for a trimeric structure with a Mr of ca. 270,000. Resorting to an anti HSP 90 probe (AC 88), we show that the native dimeric HSP 90 possess two accessible epitopes for this monoclonal antibody, while when bound to the steroid-binding subunit, only one epitope remains accessible. These data clearly suggest that the untransformed rat glucocorticoid receptor is an asymmetrical hetero-oligomeric complex.
Lefebvre
P
P
Laboratoire de Biochimie Structurale, Faculté de Médecine, Lille, France.
Sablonniere
B
B
Tbarka
N
N
Formstecher
P
P
Dautrevaux
M
M
D000818:Animals; D000911:Antibodies, Monoclonal; D002499:Centrifugation, Density Gradient; D002851:Chromatography, High Pressure Liquid; D003432:Cross-Linking Reagents; D003600:Cytosol / Q000032:analysis; D006360:Heat-Shock Proteins / Q000276:immunology / Q000302:isolation & purification*; D046911:Macromolecular Substances; D008970:Molecular Weight; D011487:Protein Conformation; D051381:Rats; D011965:Receptors, Glucocorticoid / Q000276:immunology / Q000302:isolation & purification*
D000911:Antibodies, Monoclonal; D003432:Cross-Linking Reagents; D006360:Heat-Shock Proteins; D046911:Macromolecular Substances; D011965:Receptors, Glucocorticoid