Ammonium sulfate precipitation of the glucocorticoid receptor from rat liver.
The transformed glucocorticoid receptor (GR) from rat liver precipitated at 30% saturation of ammonium sulfate and sedimented at 4.3 S on glycerol gradient centrifugation, whereas the nontransformed GR precipitated at higher concentrations of ammonium sulfate (40-50% saturation) and sedimented at 8.6 S on a gradient. Sodium dodecyl sulfate polyacrylamide gel electrophoresis showed that heat shock protein 90 (hsp 90) precipitated at 40-50% saturation of ammonium sulfate. Moreover, hsp 90 and the nontransformed GR were eluted from DEAE high performance ion-exchange chromatography at similar salt concentrations (0.22-0.23 M NaCl), whereas the transformed GR was eluted at 0.1 M NaCl. Therefore, hsp 90 seems to be responsible for the surface charge characteristics of the nontransformed GR.
Department of Biochemistry, Iwate Medical University School of Dentistry, Japan.
T
T
Nemoto
Y
Y
Ohara-Nemoto
R
R
Kurokawa
M
M
Sato
M
M
Ota
D000645:Ammonium Sulfate; D000818:Animals; D011232:Chemical Precipitation; D003600:Cytosol; D004591:Electrophoresis, Polyacrylamide Gel; D008099:Liver / Q000378:metabolism*; D008297:Male; D008970:Molecular Weight; D051381:Rats; D011919:Rats, Inbred Strains; D011965:Receptors, Glucocorticoid / Q000302:isolation & purification* / Q000378:metabolism
D011965:Receptors, Glucocorticoid; D000645:Ammonium Sulfate