Intermolecular engagement of estrogen receptors indicated by the formation of a high molecular weight complex during activation.
After exposure to ligand at 0-4 degrees C, estrogen receptors from mouse uteri characteristically eluted between thyroglobulin (Mr 669,000) and ferritin (Mr 443,000) during size-exclusion HPLC. However, when preparations were warmed with ligand under mild activating conditions, most or all of the receptor was observed as a much larger complex, which eluted between dextran blue 2000 and thyroglobulin. Formation of the large complex required ligand, was inhibited by molybdate, and occurred even in 0.4 M KCl. Slower ligand dissociation characterized the large complex, indicating that activated receptors were included preferentially. This large complex did not form when charged cytosols were aged, concentrated, or precipitated, indicating that formation was not the result of random aggregation. After exposure to conditions commonly used for activation (25 degrees C, 60 min), most receptor existed as a very large, monodisperse complex of finite size, predicting an ordered structure for these large complexes that should be useful for defining the types of proteins which can interact with estrogen receptors. Formation of the large complex was not impeded or disrupted by EDTA, RNase, DNase I, thiourea, or mercaptoethanol; however, the capacity to form this large complex was not demonstrated by preparations that had been exposed to trypsin or by the small receptor forms obtained after salt extraction. Proteolytic sensitivity and lack of sensitivity to RNase or DNase indicate that interactions between receptors and other proteins are involved in peak A formation.(ABSTRACT TRUNCATED AT 250 WORDS)
Nelson
K
K
Department of Obstetrics and Gynecology, University of Kentucky College of Medicine, Lexington 40536.
Baranowska-Kortylewicz
J
J
van Nagell
J R
JR
Gallion
H H
HH
Donaldson
E S
ES
Kenady
D E
DE
McRoberts
W P
WP
Pavlik
E J
EJ
D000818:Animals; D003600:Cytosol / Q000187:drug effects; D003850:Deoxyribonuclease I / Q000494:pharmacology; D004492:Edetic Acid / Q000494:pharmacology; D005260:Female; D006801:Humans; D008623:Mercaptoethanol / Q000494:pharmacology; D051379:Mice; D008970:Molecular Weight; D011960:Receptors, Estrogen / Q000187:drug effects / Q000378:metabolism*; D012260:Ribonucleases / Q000494:pharmacology; D013890:Thiourea / Q000494:pharmacology; D014599:Uterus / Q000187:drug effects
D011960:Receptors, Estrogen; D008623:Mercaptoethanol; D004492:Edetic Acid; D012260:Ribonucleases; D003850:Deoxyribonuclease I; D013890:Thiourea