Studies on the arrangement of glucocorticoid receptors in the plasma membrane of S-49 lymphoma cells

摘要: Abstract The presence of glucocorticoid receptors is required for glucocorticoid-mediated lymphocytolysis to take place. However, the explicit mechanism involvement this receptor continues be debated. We have recently presented evidence that response mediated by a specialised form resides in plasma membrane (mGR). Using sequential cell separation techniques (“immunopanning,” fluorescent sorting, and soft agar cloning), resultant population receptor-enriched cells remained stable provided material further analysis. mGR patching capping phenomenon originally observed with fluoresceinated monoclonal antibody was verified here electron micrographic analysis using colloidal gold-conjugated antibody. 3H-labeled antibody, radioimmunoassay developed. Trypsin treatment removed antigenic site from surface cells. Peptide mapping purified membranes reveals several trypsin α-chymotrypsin cleavage sites. Larger fragments resulted enriched versus those found depleted form, although most resulting are shared two forms. Confirmation previous studies correlating sensitivity now extended include elimination effect receptor-stripped (trypsinized) S-49 (Steroids 56:411–419, 1991)