The steroid binding domain of porcine estrogen receptor.
For the purpose of characterizing the estrogen binding domain of porcine estrogen receptor (ER), we have made use of affinity labeling of partially purified ER with [3H]tamoxifen aziridine. The labeling is very efficient and selective particularly after partial purification of ER. A 65,000-dalton (65-kDa) band was detected on the fluorogram of a sodium dodecyl sulfate-polyacrylamide gel, together with a 50-kDa band and a few more smaller bands. The 50-kDa protein appears to be a degradation product of the 65-kDa protein in view of the similar peptide map. ER was affinity labeled before or after controlled limited proteolysis with either trypsin, papain, or alpha-chymotrypsin. The labeling patterns of limited digests indicate that a fragment of about 30 kDa is relatively resistant to proteases and has a full and specific binding activity to estrogen, whereas smaller fragments have lost much of the binding activity. This fragment is very hydrophobic and probably corresponds to the carboxy half of ER.
Koike
S
S
Nii
A
A
Sakai
M
M
Muramatsu
M
M
D000818:Animals; D001665:Binding Sites; D005260:Female; D010446:Peptide Fragments / Q000032:analysis; D010447:Peptide Hydrolases; D011960:Receptors, Estrogen / Q000302:isolation & purification / Q000378:metabolism*; D013552:Swine; D013629:Tamoxifen / Q000031:analogs & derivatives* / Q000378:metabolism; D014599:Uterus / Q000378:metabolism
D010446:Peptide Fragments; D011960:Receptors, Estrogen; D013629:Tamoxifen; C031859:tamoxifen aziridine; D010447:Peptide Hydrolases