Partial purification of estradiol receptor from Xenopus laevis liver and levels of receptor in relation to estradiol concentration.
We have used ammonium sulphate precipitation followed by affinity chromatography to partially purify the estrogen receptor from Xenopus laevis liver which may control the genes for vitellogenin, the precursor of the egg yolk proteins. The rate at which receptor binds estradiol explains the kinetics of the induction of vitellogenin synthesis by estradiol, and the dissociation constant (0.5 X 10(-9) M) explains the concentration dependence of the response, which has a threshold of 10(-9) M estradiol, when 67% of the receptor is bound to estradiol. The estradiol concentration in male liver, which does not make vitellogenin, is 0.18 X 10(-9) M, sufficient to saturate 26% of the receptor, while in female liver, which makes vitellogenin continuously, the estradiol concentration is 3.5 X 10(-9) M, giving 88% saturation of receptor, suggesting that the proportion of occupied receptor decides whether or not the vitellogenin genes are active. In the physiological concentration range, estradiol modulates the level of receptor, which varies between 100 binding sites per nucleus in males and 440 in females, but artificially high concentrations of estradiol raise the level to approximately 1000 sites per nucleus. This suggests that the small increase in vitellogenin mRNA induced by physiological concentrations of estradiol is due to pre-existing receptor and that the much larger increases induced by very high concentrations depends on newly-synthesized receptor.
D000818:Animals; D002846:Chromatography, Affinity; D004958:Estradiol / Q000378:metabolism*; D005260:Female; D008099:Liver / Q000378:metabolism*; D008297:Male; D011959:Receptors, Estradiol; D011960:Receptors, Estrogen / Q000302:isolation & purification* / Q000378:metabolism; D012737:Sex Factors; D014819:Vitellogenins / Q000096:biosynthesis; D014982:Xenopus laevis
D011959:Receptors, Estradiol; D011960:Receptors, Estrogen; D014819:Vitellogenins; D004958:Estradiol