Purification of the molybdate-stabilized 9-10 S estradiol receptor from calf uterus

V. Atrache, T. Ratajczak, Sahidan Senafi, R. Hahnel

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

The molybdate-stabilized calf uterine estradiol receptor has been purified to near-homogeneity by a three-step procedure. Initial purification by heparin-Sepharose chromatography provides a concentrated receptor extract in 40% yield with a 5- to 10-fold increase in purity. The inclusion of molybdate in phosphate-buffered cytosol enhances 9-10 S receptor stability in high salt and allows elution of the oligomeric receptor complex from heparin-Sepharose with 0.4 M KCl. A second affinity step utilizing estrone carboxymethyloxime coupled to diaminoethyl bis(2-hydroxypropoxy)butane-Sepharose Cl-4B increases purification by a further 1600-fold. High performance liquid chromatography gives homogeneous receptor which migrates on sodium dodecyl sulfate-polyacrylamide gel electrophoresis as a polypeptide of M(r) ≃ 89,000. The purified molybdate-stabilized receptor sediments at 9.3 ± 0.2 S (n = 4) in glycerol gradients and has a Stokes radius of 74 ± 3 Å (n = 2) giving a calculated M(r) ≃ 290,000. These properties and the steroid-binding specificity of the purified receptor bear a close similarity to those found for the 9-10 S receptor in crude cytosol.

Original languageEnglish
Pages (from-to)5936-5941
Number of pages6
JournalJournal of Biological Chemistry
Volume260
Issue number10
Publication statusPublished - 1985
Externally publishedYes

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Estradiol Receptors
Uterus
Purification
Cytosol
Agarose Chromatography
Estrone
High performance liquid chromatography
Chromatography
Electrophoresis
Sodium Dodecyl Sulfate
Sepharose
Glycerol
Polyacrylamide Gel Electrophoresis
Sediments
Salts
Steroids
Phosphates
High Pressure Liquid Chromatography
Peptides
molybdate

ASJC Scopus subject areas

  • Biochemistry

Cite this

Purification of the molybdate-stabilized 9-10 S estradiol receptor from calf uterus. / Atrache, V.; Ratajczak, T.; Senafi, Sahidan; Hahnel, R.

In: Journal of Biological Chemistry, Vol. 260, No. 10, 1985, p. 5936-5941.

Research output: Contribution to journalArticle

Atrache, V. ; Ratajczak, T. ; Senafi, Sahidan ; Hahnel, R. / Purification of the molybdate-stabilized 9-10 S estradiol receptor from calf uterus. In: Journal of Biological Chemistry. 1985 ; Vol. 260, No. 10. pp. 5936-5941.
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