Interactions of antiglucocorticoids with glucocorticoid receptors

Six known or potential antiglucocorticoids were used as steroid probes in a competitive binding assay to elucidate the binding specificity of the agonist sites of four different rat glucocorticoid receptors. The object was to determine whether the binding of this class of steroids was particularly sensitive to subtle differences between these sites. Cytosolic extracts of heart, pancreas, kidney and liver were evaluated. The order of competitive potency for the first three preparations was found to be medroxyprogesterone greater than deoxycorticosterone greater than progesterone greater than R-5020 greater than cortexolone greater than 17-hydroxyprogesterone. The order for the glucocorticoid receptor of liver, on the other hand, was R-5020 greater than progesterone greater than deoxycorticosterone greater than medroxyprogesterone greater than cortexolone greater than 17-hydroxyprogesterone. Although this partial reversal of specificity could reflect a difference in the agonist site of the liver receptor, mixing experiments, in which cytosolic extracts of liver were incubated with kidney cytosol extracts, demonstrated that the liver cytosol contained an additional factor that could change the apparent specificity of the kidney glucocorticoid receptor. This factor was stable at 0 degrees C for at least 18 h, heat-labile and non-dialyzable. These results suggest that the liver receptor's specificity may actually be the same as the other three, but appears to be different in this type of assay because of this additional factor. It is similar that their agonist sites are most likely identical.