title: Bacterial expression, purification, and crystallization of tyrosine phosphorylated STAT proteins.
abstract: Signal Transducer and Activator of Transcription (STAT) proteins are latent cytoplasmic transcription -factors that become activated by phosphorylation at a C-terminal tyrosine residue. Upon activation STAT proteins translocate to the nucleus and bind to their specific target sites. Here, we describe the recombinant expression of tyrosine phosphorylated STAT proteins in bacteria. This method allows the production of large amounts of activated STAT proteins for structural and biochemical studies including the high-throughput screening of chemical libraries.
authors.affiliation: European Molecular Biology Laboratory, Heidelberg, Germany.
authors.forename: Florence
authors.initials: F
authors.lastname: Baudin
authors.forename: Christoph W
authors.initials: CW
authors.lastname: Müller
mesh_terms: D002852:Chromatography, Ion Exchange; D003460:Crystallization / Q000379:methods*; D004247:DNA / Q000737:chemistry / Q000378:metabolism; D024202:Electrophoretic Mobility Shift Assay; D004926:Escherichia coli / Q000235:genetics*; D015870:Gene Expression; D005818:Genetic Engineering / Q000379:methods*; D008958:Models, Molecular; D009838:Oligodeoxyribonucleotides / Q000737:chemistry / Q000378:metabolism; D010750:Phosphoproteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D010766:Phosphorylation; D010957:Plasmids / Q000235:genetics; D011487:Protein Conformation; D055503:Protein Multimerization; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D050791:STAT Transcription Factors / Q000737:chemistry* / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D014443:Tyrosine / Q000378:metabolism*
chemical_list: D009838:Oligodeoxyribonucleotides; D010750:Phosphoproteins; D011994:Recombinant Proteins; D050791:STAT Transcription Factors; D014443:Tyrosine; D004247:DNA