Purification of human recombinant GATA-1 from bacteria: implication for protein-protein interaction studies.
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.
Doubeikovskaia
Z
Z
Institute Biochemistry and Physiology of Microorganisms, Russian Academy of Sciences, Prospekt Nauki, 5, Pushchino Moscow Region, 142290, Russia. douba@rambler.ru
Aries
A
A
Jeannesson
P
P
Morle
F
F
Doubeikovski
A
A
D000818:Animals; D000918:Antibody Specificity; D001665:Binding Sites; D015153:Blotting, Western; D004268:DNA-Binding Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D050981:Erythroid-Specific DNA-Binding Factors; D004926:Escherichia coli / Q000235:genetics*; D050982:GATA1 Transcription Factor; D015870:Gene Expression; D006801:Humans; D002479:Inclusion Bodies / Q000378:metabolism; D004915:Leukemia, Erythroblastic, Acute / Q000378:metabolism; D051379:Mice; D011485:Protein Binding; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D012995:Solubility; D014157:Transcription Factors / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D014162:Transfection; D014170:Transformation, Genetic; D014407:Tumor Cells, Cultured
D004268:DNA-Binding Proteins; D050981:Erythroid-Specific DNA-Binding Factors; D050982:GATA1 Transcription Factor; C494688:GATA1 protein, human; C494689:Gata1 protein, mouse; D011993:Recombinant Fusion Proteins; D014157:Transcription Factors