Immunochemical characterization of wild-type and variant glucocorticoid receptors by monoclonal antibodies.

Open Access
Publisher Website
Google Scholar
Abstract
Monoclonal antibodies raised against the rat liver glucocorticoid receptor were used to investigate receptors of wild‐type and glucocorticoid‐resistant variants of mouse lymphoma cells. Two of the variant types contained receptors of ‘nuclear transfer deficient’ (nt‐) and ‘increased nuclear transfer’ (nti) phenotypes, respectively, while the third was of the ‘receptorless’ (r‐) phenotype with negligible hormone binding activity. Three monoclonal antibodies of the IgM class and one of the IgG class reacted with both wild‐type and nt‐ receptors but not with the steroid binding form of nti receptors. Some of the antibodies bound the wild‐type and nt‐ receptors more efficiently after activation at 20 degrees C. By use of an immuno‐competition assay we were able to detect cross‐reacting material in considerable amounts in extracts of nti and r‐ cell variants. This material was further characterized by gel filtration and immunoblotting. The immunoreactive material of wild‐type, nti and r‐ cells gave a major band of mol. wt. 94 000 upon SDS‐gel electrophoresis while the steroid‐binding polypeptides of wild‐type and nti receptors have mol. wts. of 94 000 and 40 000, respectively. The data show that in S49.1 mouse lymphoma cells the products of two receptor alleles can be distinguished.