Chemical crosslinking: a useful tool for evaluations of steroid receptor structures and their functional states in intact cells.
A procedure of chemical crosslinking of intact cells with glutaraldehyde was employed to contribute to the understanding of glucocorticoid receptor structures and their functional states in vivo. Under optimal experimental conditions, glucocorticoid binding sites were found almost equally distributed between cytosolic and nuclear fractions of crosslinked cells. Sedimentation properties of crosslinked receptor complexes in cytosolic and nuclear extracts revealed that these entities were oligomers, which heterogeneously sedimented between 11 and 4S in the presence of 0.3 M NaCl. By anion exchange chromatography, we could establish that these receptor complex oligomers behaved as untransformed forms.
Rossini
G P
GP
Istituto di Chimica Biologica, Università di Modena, Italy.
Masci
G
G
D000447:Aldehydes / Q000494:pharmacology*; D002467:Cell Nucleus / Q000378:metabolism; D002499:Centrifugation, Density Gradient; D003432:Cross-Linking Reagents; D003600:Cytosol / Q000378:metabolism; D003907:Dexamethasone / Q000378:metabolism; D005976:Glutaral / Q000494:pharmacology*; D006367:HeLa Cells / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D011965:Receptors, Glucocorticoid / Q000187:drug effects / Q000302:isolation & purification / Q000378:metabolism*; D011987:Receptors, Steroid / Q000378:metabolism*
D000447:Aldehydes; D003432:Cross-Linking Reagents; D011965:Receptors, Glucocorticoid; D011987:Receptors, Steroid; D003907:Dexamethasone; D005976:Glutaral