Absence of hydrocortisone from cytoplasmic hormone-protein complexes formed in vivo after administration of biologically active doses of [3H]hydrocortisone.
After administration of [3H]hydrocortisone to adrenalectomized rats, hormone-protein complexes were isolated from liver cytosol by DEAE-cellulose chromatography. After application of biologically active and inactive doses of hydrocortisone five binding components were detected eluting at the same salt concentrations as the hormone-protein complexes observed after incubation of cytosol with [3H]hydrocortisone in vitro. The isolated hormone-protein fractions were acidified and extracted with ethylacetate and the steroids were analyzed by thin-layer chromatography. No significant amount of hydrocortisone could be detected in any of the complexes formed in vivo 5-60 min after administration of biologically active doses of hydrocortisone. 3 xi, 11 beta, 17 alpha, 20 xi, 21-Pentahydroxypregnane, steroidal carboxy acids, glucuronides and a very polar conjugate of hydrocortisone were found in the different fractions. After an in vivo dose of hydrocortisone of about 1/5000th of the minimal dose required for enzyme induction, hydrocortisone could be found in all the cytoplasmic hormone-protein complexes formed. In contrast to the cytoplasmic hormone-protein complexes, hydrocortisone could be readily demonstrated in nuclei isolated after the administration of biologically active doses of hormone, although acid metabolites were found to represent the main part of the radioactive compounds present in the nuclei. These acid metabolites were located in the nuclear envelope. These results seem to contradict the generally accepted theory that hydrocortisone induces biosynthesis of proteins via a cytoplasmic hydrocortisone-receptor complex: after administration of biologically active doses of hydrocortisone, no such complex could be detected.
D000315:Adrenalectomy; D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism; D002855:Chromatography, Thin Layer; D003600:Cytosol / Q000378:metabolism; D006854:Hydrocortisone / Q000378:metabolism*; D008099:Liver / Q000378:metabolism*; D008297:Male; D011485:Protein Binding; D011506:Proteins / Q000378:metabolism*; D051381:Rats; D014316:Tritium
D011506:Proteins; D014316:Tritium; D006854:Hydrocortisone