Membrane Glucocorticoid Receptor Activation Induces Proteomic Changes Aligning with Classical Glucocorticoid Effects

摘要: Glucocorticoids exert rapid nongenomic effects by several mechanisms including the activation of a membrane-bound glucocorticoid receptor (mGR). Here, we report first proteomic study on mGR BSA-conjugated cortisol (Cort-BSA). A subset target proteins in data set was validated Western blot and found them responding to three additional cell lines, indicating conserved effect cells originating from different tissues. Changes proteome treated CCRF-CEM leukemia were associated with early pro-apoptotic, immune-modulatory metabolic aligning possibly “priming” classical activities cytosolic (cGR). PCR arrays investigating genes major signaling pathways indicated that does not its through transcriptional activity any most common kinases these leukemic cells, but RhoA emerged our pathway analysis. All lines tested displayed very low levels their surface. Highly sensitive specific situ proximity ligation assay visualized numbers even previously thought be negative. We obtained similar results when using distinct anti-GR monoclonal antibodies directed against N-terminal half cGR. This strongly suggests cGR have high sequence homology probably originate same gene. Furthermore, appears reside caveolae association caveolin-1 (Cav-1) clearly detected two four investigated double recognition assay. Our indicate however Cav-1 is necessary for membrane localization GR since Jurkat functional mGR, did express this caveolar protein. However, if expressed, protein dimerizes modulating function.