The subcellular distribution of glucocorticoid-receptor complexes as studied by chemical crosslinking of intact HTC cells.
Treatment of intact HTC cells with glutaraldehyde results in redistribution of glucocorticoid binding sites between cytosolic and nuclear fractions. The decrease in cytosolic receptors and their accumulation at the nuclear level were found to be directly related to the glutaraldehyde concentrations employed in our procedure and inversely related to the cell density of samples. When the data from eleven separate experiments were combined, and analyzed by linear regression of cytosolic and nuclear levels of receptor complexes vs the ratios between the DNA and glutaraldehyde concentration of our samples, two lines were obtained whose intercepts on the ordinate yielded values of cytosolic and nuclear receptors corresponding to 37.5 and 62.5% of the total cellular pool, respectively. When we compared the subcellular redistribution of glucocorticoid receptor to that of the cytosolic enzyme lactate dehydrogenase upon HTC cell crosslinking with glutaraldehyde, we found that the cytosolic and nuclear levels of the enzyme were 53.2 and 46.8% of the total content, respectively. If the subcellular distribution of glucocorticoid receptor is corrected for the artefactual redistribution induced by crosslinking, using the values obtained for lactate dehydrogenase, it can be concluded that glucocorticoid receptors in HTC cells are distributed between cytosol and nuclei in a ratio which is about 2:1. Our findings lend further support to the conclusion that only a portion of glucocorticoid receptor is cytosolic in intact cells.
Rossini
G P
GP
Istituto di Chimica Biologica, Università di Modena, Italy.
Malaguti
C
C
D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D003432:Cross-Linking Reagents; D003600:Cytosol / Q000378:metabolism; D005976:Glutaral / Q000494:pharmacology; D006367:HeLa Cells; D006801:Humans; D011965:Receptors, Glucocorticoid / Q000378:metabolism*; D013347:Subcellular Fractions
D003432:Cross-Linking Reagents; D011965:Receptors, Glucocorticoid; D005976:Glutaral