Transformation of the glucocorticoid-receptor complex by heating the cytosol in the presence of calcium is accompanied by formation of a series of truncated complexes, of which DI and DIIc are the major members. Formation of DIIc (but not of DI) is inhibited by leupeptin, and the intact transformed complex DIIa appears instead. Estimation of the molecular weights and Stokes' radii of all major complexes revealed that forms DI and DIIc have the same Mr, 48 kDa, but differ in shape, and appear to be digestion products generated by cleavage at the same site. Proteolysis of glucocorticoid receptor, covalently labelled with [3H]dexamethasone mesylate in rat thymus and brain cytosol, corroborated these findings and further implied that DI is the product of digestion of the non-transformed form of the receptor. Covalently labelled receptor fragments, related to the products formed when cytosol is heated, are detected in the nuclei of thymocytes, implying that the same proteolytic cleavages sites are involved in receptor turnover. Cleavage sites in the non-transformed covalently labelled receptor were identified in the "stepladder" of fragments of Mr, 85, 65, 49, 35, 27-30 kDa, generated in the absence of calcium, with an additional 78 kDa fragment in its presence. In the transformed conformation, two of the cleavage sites giving rise to the 65 and 35 kDa fragments, appear to be protected. It is speculated that the change in the proteolytic susceptibility of the cleavage site for the 35 kDa fragment relates to the "unmasking" of enhancer-activating and/or DNA-binding receptor functions previously postulated.