Purification and characterization of mouse uterine estrogen receptor under conditions of varying hormonal status.
Mouse uterine estrogen receptor (ER) was purified about 11,000-fold from normal mouse uteri by affinity chromatography. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the purified ER demonstrated a major component of 65,000 mol wt with minor fragments of the 54,000- and 37,000-dalton species, as judged by affinity labeling with [3H]tamoxifen aziridine and immunodetection with an ER monoclonal antibody (H-222). The minor fragments were not detected with additional monoclonals (H-226 or D-547), which recognize different domains on the ER molecule. Two-dimensional gel electrophoresis revealed that the major component with a mol wt of 65,000 had a pI of about 6.5. The 54,000- and 37,000-dalton components had similar pI values. Saturation binding and Scatchard plot analysis of purified ER yielded one class of binding sites with an apparent dissociation constant of about 1.4 nM. Changes in the hormonal status resulted in changes in the size of the ER even in the presence of molybdate and leupeptin.
Horigome
T
T
Laboratory of Reproductive and Developmental Toxicology, National Institute of Environmental Health Sciences, Research Triangle Park, North Carolina 27709.
Golding
T S
TS
Quarmby
V E
VE
Lubahn
D B
DB
McCarty
K
K
Korach
K S
KS
D000818:Animals; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D004958:Estradiol / Q000378:metabolism; D005260:Female; D051379:Mice; D008813:Mice, Inbred ICR; D008970:Molecular Weight; D010052:Ovariectomy; D011960:Receptors, Estrogen / Q000302:isolation & purification / Q000378:metabolism*; D012016:Reference Values; D013629:Tamoxifen / Q000031:analogs & derivatives / Q000378:metabolism; D014599:Uterus / Q000378:metabolism*
D011960:Receptors, Estrogen; D013629:Tamoxifen; D004958:Estradiol; C031859:tamoxifen aziridine