Identification of hormone-interacting amino acid residues within the steroid-binding domain of the glucocorticoid receptor in relation to other steroid hormone receptors.

Purified rat liver glucocorticoid receptor was covalently charged with 3Hglucocorticoid by photoaffinity labeling (UV irradiation of 3Htriamcinolone acetonide-glucocorticoid receptor) or affinity labeling (incubation with 3Hdexamethasone mesylate). After labeling, separate samples of the denatured receptor were cleaved with trypsin (directly or after prior succinylation), chymotrypsin, and cyanogen bromide. Labeled residues in the peptides obtained were identified by radiosequence analysis. The peaks of radioactivity corresponded to Met-622 and Cys-754 after photoaffinity labeling with 3Htriamcinolone acetonide and Cys-656 after affinity labeling with 3Hdexamethasone mesylate. The labeled residues are all positioned within hydrophobic segments of the steroid-binding domain. The patterns of hydropathy and secondary structure for the glucocorticoid receptor are highly similar to those for the progestin receptor and similar but less so to those for the estrogen receptor and to those for c-erb A.