PMID: 444462May 29, 1979Paper

Estrogen-binding proteins of calf uterus. Purification to homogeneity of receptor from cytosol by affinity chromatography

Biochemistry
V Sica, F Bresciani

Abstract

The estrogen receptor has been purified to homogeneity from calf uterus cytosol by sequential affinity chromatography by using heparin--Sepharose 4B and 17-hemisuccinyl-17beta-estradiol-ovalbumin--Sepharose 4B. The procedure yields about 1.2 mg of receptor protein from 1 kg of calf uteri, with a recovery of 53%. The receptor protein, as a complex with 17beta-[3H]estradiol, is purified more than 99%. A single band is seen on polyacrylamide gel ectrophoresis under nondenaturing conditions. 17beta-[3H]Estradiol comigrates with the protein band. As computed from the specific activity of radioactive hormone, 64,450 g of purified receptor protein binds 1 mol of 17beta-estradiol. 17beta-[3H]Estradiol bound to the protein is displaced by estrogenic steriods but not by progesterone, testosterone, or cortisone. As judged by chromatography on calibrated Sephadex G-200 columns, the purified receptor is identical with native receptor in crude cytosol: both show a Stokes radius of 6.4 nm. On sucrose gradient in low-salt buffer, the purified receptor sediments at 8 S. On electrophoresis in NaDodSO4 gels, the purified receptor migrates as a single protein band with an apparent molecular weight of 70,000. The sedimentation coefficient measured ...Continue Reading

References

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Jun 1, 1975·Proceedings of the National Academy of Sciences of the United States of America·A A WaldmanJ Goldstein
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Oct 1, 1962·Analytical Biochemistry·T BITTER, H M MUIR

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Citations

Jan 1, 1981·Breast Cancer Research and Treatment·L P PertschukN D Bloom
Nov 14, 1979·Biochemical and Biophysical Research Communications·R N Taylor, R G Smith
Jan 1, 1980·Journal of Steroid Biochemistry·G A PucaV Sica
Aug 1, 1981·Journal of Steroid Biochemistry·P LustenbergerM Dautrevaux
Dec 1, 1981·Journal of Steroid Biochemistry·T C Madhok, B S Leung
Mar 1, 1982·Journal of Steroid Biochemistry·G L Greene, E V Jensen
Jun 1, 1984·Journal of Steroid Biochemistry·W A McBlain, G Shyamala
Jan 1, 1986·Journal of Steroid Biochemistry·G L Greene, M F Press
Mar 1, 1986·Journal of Steroid Biochemistry·A D LinstedtR M Brenner
Dec 1, 1988·Journal of Steroid Biochemistry·T RatajczakR Hähnel
Jun 1, 1983·Molecular and Cellular Endocrinology·L H SunF R Frankel
Nov 17, 1980·Biochimica Et Biophysica Acta·W Schneider, M Gschwendt
Jan 1, 1986·Meat Science·M MattioliF Chiesa
Sep 1, 1980·Proceedings of the National Academy of Sciences of the United States of America·G L GreeneE V Jensen
May 1, 1980·European Journal of Biochemistry·H M Westphal, M Beato
Jun 26, 1985·Journal of Chromatography·S M HyderJ L Wittliff
Feb 22, 1983·Biochimica Et Biophysica Acta·J R Hubbard, M Kalimi
Feb 1, 1984·Analytical Biochemistry·T R Van OosbreeG C Mueller
Jan 1, 1984·Annals of the New York Academy of Sciences·D J TindallD R Rowley
Mar 1, 1983·Clinics in Endocrinology and Metabolism·K S McCartyK S McCarty
Dec 1, 1980·Analytical Biochemistry·P K SmithG T Hermanson
May 31, 1985·Biochemical and Biophysical Research Communications·S CollinsM A Marletta
Jan 1, 1984·Acta Radiologica. Oncology·H S PoulsenN Brünner
Jan 1, 1983·Journal of Receptor Research·T R Van OosbreeG C Mueller

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