Effect of chemical inactivating agents on glucocorticoid receptor proteins in mouse and hamster cells.
The ffect of N-ethylmaleimide and iodoacetamide on the glucocorticoid receptor activity extracted from the cytosol of either mouse of hamster cells has been investigated. Treatment of mouse or hamster cytosol with N-ethylmaleimide or iodoacetamide rapidly inactivates the [3H]glucocorticoid hormone binding activity of either cytosol. Prebinding the glucocorticoid hormone, dexamethasone, to the cytosol receptor blocks the rapid inactivation of the receptor by N-ethylmaleimide. Treatment of the prebound hormone-receptor complex with iodoacetamide prevents the subsequent binding of the hormone-receptor complex to DNA without causing a dissociation of the complex. Although the conclusions may be limited by the lack of purity of the receptor, the results suggest that a sulfhydryl group is involved in the binding of glucocorticoid hormones to the receptor protein. In addition, the results suggest that iodoacetamide is inactivating a separate chemical site which is necessary for the binding of the hormone-receptor complex to DNA.
D001665:Binding Sites; D002118:Calcium / Q000494:pharmacology; D002460:Cell Line; D003600:Cytosol / Q000378:metabolism; D003907:Dexamethasone / Q000378:metabolism*; D004229:Dithiothreitol / Q000494:pharmacology; D005033:Ethylmaleimide / Q000494:pharmacology*; D005938:Glucocorticoids / Q000494:pharmacology*; D007460:Iodoacetamide / Q000494:pharmacology*; D007461:Iodoacetates / Q000494:pharmacology*; D007700:Kinetics; D008274:Magnesium / Q000494:pharmacology; D008345:Manganese / Q000494:pharmacology; D011485:Protein Binding; D011956:Receptors, Cell Surface / Q000187:drug effects*; D013696:Temperature
D005938:Glucocorticoids; D007461:Iodoacetates; D011956:Receptors, Cell Surface; D008345:Manganese; D003907:Dexamethasone; D008274:Magnesium; D005033:Ethylmaleimide; D002118:Calcium; D004229:Dithiothreitol; D007460:Iodoacetamide