Purification and characterization of a nonhormone-binding component of the nontransformed glucocorticoid receptor from rat liver.
The nontransformed glucocorticoid receptor (GR) and an 88-kDa protein in rat liver cytosol were selectively adsorbed on protamine- and arginine-Sepharose. The 88-kDa protein was purified from rat liver cytosol to homogeneity by precipitation with protamine sulfate, followed by DEAE-ion exchange chromatography, gel chromatography, and DEAE ion-exchange high-performance liquid chromatography. The 88-kDa protein appeared to be present as a dimer at both low and high salt concentrations. The physicochemical properties and the amino acid composition of the 88-kDa protein were almost the same as those of heat-shock protein 90 of HeLa cells and yeast. Preincubation of the GR with the polyclonal antibody raised against the 88-kDa protein increased the sedimentation coefficient of the nontransformed GR, but did not change that of the transformed GR. These results indicate that heat-shock protein 90 is associated with rat liver nontransformed GR and is responsible for the interaction of GR with protamine.
Department of Biochemistry, Iwate Medical University School of Dentistry.
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Nemoto
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Ohara-Nemoto
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Ota
D000315:Adrenalectomy; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002021:Buffers; D002498:Centrifugation; D003600:Cytosol / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D005990:Glycerol; D008099:Liver / Q000378:metabolism*; D008297:Male; D051381:Rats; D011919:Rats, Inbred Strains; D011965:Receptors, Glucocorticoid / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D012967:Sodium Dodecyl Sulfate
D000596:Amino Acids; D002021:Buffers; D011965:Receptors, Glucocorticoid; D012967:Sodium Dodecyl Sulfate; D005990:Glycerol