Localization of pyridoxal phosphate binding site on the mero-receptor domain of the glucocorticoid receptor.
Previous studies have demonstrated that the vitamin pyridoxal phosphate can alter the physicochemical properties of glucocorticoid receptors. We now report the localization of a pyridoxal phosphate binding site within the mero-receptor domain of this glucocorticoid receptor. Mero-glucocorticoid receptors that are generated by trypsin (10 micrograms/ml) or chymotrypsin (100 micrograms/ml) digestion of intact receptors sediment as 2.6 S species on 5-20% sucrose gradients in the presence or absence of pyridoxal phosphate. Mero-glucocorticoid receptors prepared by exogenous proteinases are hydrophobic and show no affinity for DEAE Bio-Gel A. Treating either trypsin-generated or chymotrypsin-generated mero-receptors with pyridoxal phosphate rapidly converts the proteins (60 and 35%, respectively) into forms that bind to DEAE Bio-Gel A. Induction of DEAE binding is specific to pyridoxal phosphate, for treating mero-receptors with pyridoxal, pyridoxamine or pyridoxine phosphate is ineffective. Furthermore, DEAE binding cannot be induced by adding other pyridoxal phosphate-treated cytosols to untreated mero-receptors. High-resolution polyacrylamide gel isoelectric focussing studies indicated that treating mero-receptor generated by either proteinase with pyridoxal phosphate shifted the isoelectric points of both to lower pH values. The conversion of the mero-receptor to a more acidic form also occurred when the intact glucocorticoid receptor was treated with the vitamin prior to proteolysis. These studies localize at least one pyridoxal phosphate binding site on the mero-receptor domain of the rat thymocyte glucocorticoid receptor.
D000818:Animals; D001665:Binding Sites; D002918:Chymotrypsin; D007526:Isoelectric Point; D008297:Male; D010446:Peptide Fragments; D011487:Protein Conformation; D011732:Pyridoxal Phosphate / Q000378:metabolism*; D051381:Rats; D011965:Receptors, Glucocorticoid / Q000378:metabolism*; D011987:Receptors, Steroid / Q000378:metabolism*; D012995:Solubility; D013950:Thymus Gland; D014357:Trypsin
D010446:Peptide Fragments; D011965:Receptors, Glucocorticoid; D011987:Receptors, Steroid; D011732:Pyridoxal Phosphate; D002918:Chymotrypsin; D014357:Trypsin