Reconstitution of the 9 S estrogen receptor with heat shock protein 90.
As a first step in the investigation of the reconstitution of steroid hormone receptor systems, we studied the reconstitution of 9 S estrogen receptor (ER) from purified vero ER, which is the estradiol binding subunit, and heat shock protein 90 (hsp 90). By using a phosphate buffer containing molybdate, thiocyanate, dimethylformamide, glycerol, etc., vero ER could be converted to 9 S ER with hsp 90, but not with the control protein, ovalbumin. Inactivation of ER during the reconstitution was suppressed partially by hsp 90, but not by ovalbumin. Like native 8 S ER, the reconstituted ER was sedimented at about 8.9 S and 4.6 S on glycerol gradient centrifugation in low and high salt buffers, respectively.
Inano
K
K
Department of Biochemistry, Faculty of Science, Niigata University, Japan.
Haino
M
M
Iwasaki
M
M
Ono
N
N
Horigome
T
T
Sugano
H
H
D000818:Animals; D002417:Cattle; D002499:Centrifugation, Density Gradient; D005260:Female; D006360:Heat-Shock Proteins / Q000502:physiology*; D011960:Receptors, Estrogen / Q000378:metabolism*
D006360:Heat-Shock Proteins; D011960:Receptors, Estrogen