Estrogen affinity crosslinking to tyrosinase-like immunoreactive proteins of rat uterine nuclear extracts.
We have previously described tyrosinase-like proteins of rat uterine nuclear extracts with type II estrogen binding characteristics. In this paper we have been able to affinity label these polypeptides with radio-iodinated estradiol. The major label at approximately 33-38 kDa comigrates with a approximately 36 kDa tyrosinase immunoreactive band assessed by autoradiograms and Western blots following electrophoresis. A minor label was also detected at approximately 45 kDa. The label is attenuated by excess quercetin hence these proteins are believed to represent putative type II estrogen binding sites that bind this bioflavonoid. These estrogen binding proteins are distinct from the estrogen receptor as judged by immunoblotting. The affinity crosslinking will be a useful approach in the purification of tyrosinase like proteins.
Garai
J
J
Department of Cell Biology, Baylor College of Medicine, Houston, TX 77030.
Clark
J H
JH
D000818:Animals; D001345:Autoradiography; D015153:Blotting, Western; D002467:Cell Nucleus / Q000737:chemistry*; D003432:Cross-Linking Reagents; D004958:Estradiol / Q000378:metabolism*; D004970:Estrone / Q000378:metabolism; D005260:Female; D006632:Histamine / Q000378:metabolism; D007457:Iodine Radioisotopes; D014442:Monophenol Monooxygenase / Q000378:metabolism*; D009687:Nuclear Proteins / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013205:Staphylococcal Protein A / Q000378:metabolism; D014599:Uterus / Q000648:ultrastructure*
D003432:Cross-Linking Reagents; D007457:Iodine Radioisotopes; D009687:Nuclear Proteins; D013205:Staphylococcal Protein A; D004970:Estrone; D004958:Estradiol; D006632:Histamine; D014442:Monophenol Monooxygenase