Maximizing the purification of the activated glucocorticoid receptor by DNA-cellulose chromatography.

The Biochemical journal ·Vol. 171 ·No. 1 ·1978-04-01 ·Pages 177-83

Eisen HJ, Glinsmann WH

Abstract

With heat treatment (20 degrees C for 30 min), the glucocorticoid-receptor complex becomes 'activated' and undergoes an increase in affinity for DNA. A two-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 first chromatographic step was critically dependent on ionic conditions and the ratio of cytosol chromatographed to DNA-cellulose. A purification of 11000-fold (85% yield) was achieved by this procedure. The partially purified receptor complex was taken up by rat liver nuclei.

MeSH Terms

Animals Cell Nucleus/metabolism Cellulose Chromatography, Affinity/methods DNA Electrophoresis, Polyacrylamide Gel Hot Temperature In Vitro Techniques Liver/metabolism Male Protein Binding Rats Receptors, Glucocorticoid/isolation & purification,metabolism Receptors, Steroid/isolation & purification Triamcinolone/metabolism

Chemicals

Receptors, Glucocorticoid Receptors, Steroid Triamcinolone Cellulose DNA

Authors & Affiliations

2 authors, click to expand affiliations / ORCID

Eisen H J

Glinsmann W H

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24 references, click to expand

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Article Info

Journal

The Biochemical journal

Abbr.

Biochem J

ISSN

0264-6021

DOI

10.1042/bj1710177

Published

1978-04-01

Pages

177-83

Language

English

Region

England

NLM ID

2984726R

PMCID

PMC1184147

Subset

External Links

PubMed source DOI: 10.1042/bj1710177 PMC Full Text View journal details

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Maximizing the purification of the activated glucocorticoid receptor by DNA-cellulose chromatography.

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