Metal binding sites of the estradiol receptor from calf uterus and their possible role in the regulation of receptor function.
The existence of putative metal binding sites on the estradiol receptor (ER) molecule from calf uterus was evaluated by immobilizing various divalent metals to iminodiacetate-Sepharose. ER from both crude and highly purified preparations binds to metal-containing adsorbents complexed with Zn(II), Ni(II), Co(II), and Cu(II), but not to those complexed with Fe(II) and Cd(II). Elution of ER was obtained by chelating agents or by imidazole, thus indicating that histidine residues on the ER molecule are involved in the interaction with the metal. Analysis of affinity-labeled ER by [3H]tamoxifen aziridine after elution from a column of Zn(II)-charged iminodiacetate-Sepharose showed that ER fragments obtained by extensive trypsinization were also bound. Zn(II) and the same other metals able to bind ER, when immobilized on resins, inhibit the binding of estradiol to the receptor at micromolar concentrations. This inhibition is noncompetitive and can be reversed by EDTA. The inhibition of the hormone binding was still present after trypsin treatment of the cytosol, and it was abolished by preincubation with the hormone. Micromolar concentrations of these metals were able to block those chemical-physical changes occurring during the process of ER transformation in vitro. Furthermore, if added to pretransformed ER-hormone complex, they strongly inhibited the binding of the complex to isolated nuclei. The presence of metal binding sites that modulate the ER activity in the hormone binding domain of ER is therefore speculated. Since progesterone receptor showed the same pattern of binding and elution from metal-containing adsorbents, the presence of metal binding regulatory sites could be a property of all steroid receptors.
Medici
N
N
Istituto di Patologia Generale ed Oncologia, III Cattedra, I Facoltà di Medicina e Chirurgia, Naples, Italy.
Minucci
S
S
Nigro
V
V
Abbondanza
C
C
Armetta
I
I
Molinari
A M
AM
Puca
G A
GA
D000818:Animals; D001665:Binding Sites; D002417:Cattle; D002467:Cell Nucleus / Q000378:metabolism; D002846:Chromatography, Affinity; D003035:Cobalt / Q000378:metabolism; D003300:Copper / Q000378:metabolism; D004958:Estradiol / Q000378:metabolism; D005260:Female; D066298:In Vitro Techniques; D008670:Metals / Q000378:metabolism*; D009532:Nickel / Q000378:metabolism; D011959:Receptors, Estradiol / Q000378:metabolism*; D014599:Uterus / Q000378:metabolism*; D015032:Zinc / Q000378:metabolism
D008670:Metals; D011959:Receptors, Estradiol; D003035:Cobalt; D004958:Estradiol; D003300:Copper; D009532:Nickel; D015032:Zinc