Purification of human recombinant GATA-1 from bacteria: implication for protein-protein interaction studies.

Protein expression and purification ·Vol. 23 ·No. 3 ·2001-12-00 ·Pages 426-31

Doubeikovskaia Z, Aries A, Jeannesson P, Morle F, Doubeikovski A

Abstract

GATA-1 is a key regulator of terminal erythroid differentiation in mammals and birds. The structural and biochemical studies of human GATA-1 (hGATA-1) are limited by the difficulty of its purification in a sufficient amount. Here we describe the procedure for obtaining pure bacterial recombinant hGATA-1 in an active functional state. We demonstrate that this protein may be successfully used for preparing an affinity column, producing GATA-1-specific rabbit polyclonal antibodies, and studying DNA-protein and protein-protein interactions.

MeSH Terms

Animals Antibody Specificity Binding Sites Blotting, Western DNA-Binding Proteins/genetics,isolation & purification,metabolism Erythroid-Specific DNA-Binding Factors Escherichia coli/genetics GATA1 Transcription Factor Gene Expression Humans Inclusion Bodies/metabolism Leukemia, Erythroblastic, Acute/metabolism Mice Protein Binding Rabbits Recombinant Fusion Proteins/chemistry,isolation & purification,metabolism Solubility Transcription Factors/genetics,isolation & purification,metabolism Transfection Transformation, Genetic Tumor Cells, Cultured

Chemicals

DNA-Binding Proteins Erythroid-Specific DNA-Binding Factors GATA1 Transcription Factor GATA1 protein, human Gata1 protein, mouse Recombinant Fusion Proteins Transcription Factors

Authors & Affiliations

5 authors, click to expand affiliations / ORCID

Doubeikovskaia Z

Institute Biochemistry and Physiology of Microorganisms, Russian Academy of Sciences, Prospekt Nauki, 5, Pushchino Moscow Region, 142290, Russia. douba@rambler.ru

Aries A

Jeannesson P

Morle F

Doubeikovski A

Article Info

Journal

Protein expression and purification

Abbr.

Protein Expr Purif

ISSN

1046-5928

DOI

10.1006/prep.2001.1533

douba@rambler.ru

Published

2001-12-00

Pages

426-31

Language

English

Region

United States

NLM ID

9101496

Subset

External Links

PubMed source DOI: 10.1006/prep.2001.1533 View journal details

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Purification of human recombinant GATA-1 from bacteria: implication for protein-protein interaction studies.

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