Reversible Denaturation of the Estrogen Receptor and Estimation of Polypeptide Chain Molecular Weight*

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Abstract
The estrogen receptor protein in rats loses its ability to bind to estrogens upon denaturation with Na dodecyl sulfate and 2-mercaptoethanol. Binding activity is recovered at 60-80% efficiency upon removal of the denaturants, equilibration with 6 M guanidine hydrochloride, and dilution into buffer containing estrogen. Renatured receptor is similar to native receptor in affinity for 17.beta.-estradiol and ability to bind DNA. Detection of receptor activity after Na dodecyl sulfate-polyacrylamide gel electrophoresis of uterine or pituitary cytosol or of uterine nuclear extracts reveals a single unique polypeptide species of 65,000 MW.