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 [3H]glucocorticoid by photoaffinity labeling (UV irradiation of [3H]triamcinolone acetonide-glucocorticoid receptor) or affinity labeling (incubation with [3H]dexamethasone 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 [3H]triamcinolone acetonide and Cys-656 after affinity labeling with [3H]dexamethasone 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.
Carlstedt-Duke
J
J
Department of Medical Nutrition, Karolinska Institutet, Huddinge University Hospital, Sweden.
Strömstedt
P E
PE
Persson
B
B
Cederlund
E
E
Gustafsson
J A
JA
Jörnvall
H
H
D000345:Affinity Labels; D000595:Amino Acid Sequence; D000818:Animals; D002918:Chymotrypsin; D003488:Cyanogen Bromide; D003907:Dexamethasone / Q000031:analogs & derivatives* / Q000378:metabolism; D008099:Liver / Q000378:metabolism*; D010446:Peptide Fragments / Q000302:isolation & purification / Q000378:metabolism; D011489:Protein Denaturation; D051381:Rats; D011965:Receptors, Glucocorticoid / Q000302:isolation & purification / Q000378:metabolism*; D011987:Receptors, Steroid / Q000378:metabolism*; D014222:Triamcinolone Acetonide / Q000378:metabolism*; D014357:Trypsin
D000345:Affinity Labels; D010446:Peptide Fragments; D011965:Receptors, Glucocorticoid; D011987:Receptors, Steroid; D003907:Dexamethasone; D002918:Chymotrypsin; D014357:Trypsin; D014222:Triamcinolone Acetonide; C031710:dexamethasone 21-methanesulfonate; D003488:Cyanogen Bromide