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- W2028908780 abstract "A new antiestrogen affinity ligand for the covalent labeling of estrogen receptors, [3H]desmethylnafoxidine aziridine, has been used to investigate the salt- and temperature-independent formation of DNA-binding estrogen receptor forms from untransformed (300 kilodaltons) receptor. Calf uterine estrogen receptor proteins labeled with [3H]estradiol or [3H]desmethylnafoxidine aziridine were quantitatively transformed (>90%) to their DNA-binding configuration in low ionic strength buffers by brief exposure to 3 m urea at 0 C. The urea effect was hormone-dependent and partially reversible. The transformed receptors were purified (ca 250-fold) by affinity chromatography on single-stranded DNAagarose in the continued presence of 3 m urea to prevent transformation reversal. Scatchard analyses revealed a single class of high affinity radioligand binding sites (Kd = 0.34 nm) unchanged by ureainduced transformation and purification. The DNAbinding receptor form labeled with [3H]desmethylnafoxidine aziridine was stable as a probable dimer in 3 m urea with 0.4 m KCI and displayed no evidence of size (Stokes radius 7.3 to 7.5 nm; 4.2 to 4.3 S; Mr = 136,800) heterogeneity. Sodium dodecyl sulfatepolyacrylamide gradient gel electrophoresis indicated the presence of an intact 67 kDa steroidbinding receptor subunit. Reverse-phase chromatography of the covalently labeled receptor on C4 and phenyl stationary phases revealed no evidence of structural heterogeneity. The surface charge of the estrogen- and antiestrogen-receptor complexes, however, was distinctly different in both the presence and absence of 3 m urea. Thus, exposure to urea was an effective salt- and temperature-independent means for achieving the complete transformation of receptor to its stable DNA-binding dimer configuration. The ligand-induced differences in receptor surface charge and the urea effects on DNAbinding (but not hormone-binding) suggest that both electrostatic and hydrophobic or hydrogen bonding receptor domains are influenced by ligand binding." @default.
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- W2028908780 date "1990-02-01" @default.
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- W2028908780 title "Unique Molecular Properties of a Urea- and Salt-Stable DNA-Binding Estrogen Receptor Dimer Covalently Labeled with the Antiestrogen [<sup>3</sup>H] Desmethylnafoxidine Aziridine. A Comparison with the Estrogen- Receptor Complex." @default.
- W2028908780 doi "https://doi.org/10.1210/mend-4-2-255" @default.
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