Maximizing the purification of the activated glucocorticoid receptor by DNA-cellulose chromatography
- 1 April 1978
- journal article
- research article
- Published by Portland Press Ltd. in Biochemical Journal
- Vol. 171 (1) , 177-183
- https://doi.org/10.1042/bj1710177
Abstract
With heat treatment (20.degree. C for 30 min), the glucocorticoid-receptor complex becomes activated and undergoes an increase in affinity for DNA. A 2-stage procedure was used to separate sequentially the rat liver glucocorticoid-receptor complex from proteins with high and low affinity for DNA. DNA-cellulose column chromatography of unheated cytosol resulted in the retention of DNA-binding proteins, but not the unactivated receptor complex. Heat treatment of the column eluate resulted in increased affinity of the receptor complex to DNA, and chromatography on DNA-cellulose then yielded receptor complex free from proteins with low affinity for DNA. Removal of DNA-binding proteins during the 1st chromatographic step was critically dependent on ionic conditions and the ratio of cytosol chromatographed to DNA-cellulose. A purification of 11,000-fold (85% yield) was achieved by this procedure. The partially purified receptor complex was taken up by rat liver nuclei.This publication has 24 references indexed in Scilit:
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