Immunochemical comparison of mutant glucocorticoid receptors and wild type receptor fragments produced by neutrophil elastase and chymotrypsin.
We characterized the glucocorticoid receptor fragments produced by neutrophil elastase and compared these receptor fragments to nuclear transfer increased (nti) mutant receptors. Neutrophil elastase and chymotrypsin digested [3H]dexamethasone 21-mesylate labeled receptors at different sites to produce 52 kDa and 42 kDa fragments respectively. Both the 52 kDa elastolytic receptor fragments and 42 kDa chymotryptic receptor fragments bound to DNA-cellulose and were immunoadsorbed by anti-glucocorticoid receptor monoclonal antibodies (BUGR2). More extensive digestion of labeled receptors by neutrophil elastase produced 29 kDa receptor fragments that did not bind to DNA-cellulose and did not react with BUGR2 antibodies. The size of nti mutant receptors from S49 mouse lymphoma cell variants is intermediate between that of the 52 kDa elastolytic receptor fragments and 42 kDa chymotryptic receptor fragments. The nti receptors bound to DNA-cellulose with the same affinity as the 52 kDa elastolytic receptor fragments. However, the nti receptors were not immunoadsorbed by BUGR2 antibodies and did not react with these antibodies on Western blot analysis of denatured cellular proteins. The results indicate that 52 kDa elastolytic receptor fragments, 42 kDa chymotryptic receptor fragments and nti mutant receptors correspond to the same region of the receptor molecule. The failure of nti receptors to react with BUGR2 antibodies suggests that the nti receptors may have an altered sequence compared to the corresponding region of normal receptors.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D002846:Chromatography, Affinity; D002918:Chymotrypsin / Q000378:metabolism*; D008223:Lymphoma / Q000201:enzymology; D051379:Mice; D009363:Neoplasm Proteins / Q000378:metabolism; D009504:Neutrophils / Q000201:enzymology*; D010196:Pancreatic Elastase / Q000378:metabolism*; D010446:Peptide Fragments / Q000276:immunology*; D011965:Receptors, Glucocorticoid / Q000235:genetics / Q000276:immunology* / Q000378:metabolism
D000911:Antibodies, Monoclonal; D009363:Neoplasm Proteins; D010446:Peptide Fragments; D011965:Receptors, Glucocorticoid; D002918:Chymotrypsin; D010196:Pancreatic Elastase