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PMID: 3316979 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Purification of the c-fos enhancer-binding protein.

Molecular and cellular biology ·Vol. 7 ·No. 10 ·1987-10-00 ·Pages 3482-9

Prywes R, Roeder RG

Abstract

We have purified the c-fos enhancer-binding protein from HeLa cell nuclear extracts. The key purification steps involved chromatography on a nonspecific DNA affinity column, from which binding activity and other protein were eluted at low salt concentrations, followed by chromatography on a specific oligonucleotide affinity column, from which the enhancer binding activity was specifically eluted at high salt concentrations. The purified protein had a strong affinity for the c-fos enhancer dyad symmetry sequence, with an equilibrium dissociation constant of 3.3 x 10(-11) M. This affinity was at least 50,000-fold stronger than that found for nonspecific DNA sequences.

MeSH Terms
Chromatography DNA-Binding Proteins/isolation & purification Deoxyribonuclease I Enhancer Elements, Genetic HeLa Cells Humans Nuclear Proteins/isolation & purification Oligodeoxyribonucleotides/metabolism Protein Binding Proto-Oncogene Proteins/genetics Transcription Factors/isolation & purification
Chemicals
DNA-Binding Proteins Nuclear Proteins Oligodeoxyribonucleotides Proto-Oncogene Proteins Transcription Factors Deoxyribonuclease I
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Prywes R
Laboratory of Biochemistry and Molecular Biology, Rockefeller University, New York, New York 10021.
Roeder R G
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.7.10.3482-3489.1987
Published
1987-10-00
Pages
3482-9
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC368000
Subset
IM
External Links
PubMed source DOI: 10.1128/mcb.7.10.3482-3489.1987 PMC Full Text View journal details
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