title: Expression and Purification of Soluble STAT5b/STAT3 Proteins for SH2 Domain Binding Assay.
abstract: When a large hydrophobic full-length protein is expressed in bacteria, it is often challenging to obtain recombinant proteins in the soluble fraction. One way to overcome this challenge is expression of deletion mutants that have improved solubility while maintaining biological activity. In this chapter, we describe a protocol for expression of truncated forms of STAT5b and STAT3 proteins that are soluble and retain SH2-mediated activity for phospho-Tyr peptide recognition.
authors.affiliation: Center for Drug Discovery, Graduate School of Pharmaceutical Sciences, University of Shizuoka, 52-1 Yada, Suruga-ku, Shizuoka, 422-8526, Japan. aasai@u-shizuoka-ken.ac.jp.
authors.forename: Akira
authors.initials: A
authors.lastname: Asai
authors.affiliation: Center for Drug Discovery, Graduate School of Pharmaceutical Sciences, University of Shizuoka, 52-1 Yada, Suruga-ku, Shizuoka, 422-8526, Japan.|Innovative Research Division, Mitsubishi Tanabe Pharma Corporation, Saitama, Japan.
authors.forename: Kazuyuki
authors.initials: K
authors.lastname: Takakuma
mesh_terms: D001681:Biological Assay; D015870:Gene Expression; D006801:Humans; D010957:Plasmids / Q000235:genetics; D011485:Protein Binding; D011994:Recombinant Proteins; D050796:STAT3 Transcription Factor / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification* / Q000378:metabolism; D050799:STAT5 Transcription Factor / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification* / Q000378:metabolism; D012995:Solubility; D018909:src Homology Domains
chemical_list: D011994:Recombinant Proteins; D050796:STAT3 Transcription Factor; D050799:STAT5 Transcription Factor
keywords: Purification; SH2 domain; STAT3; STAT5b; Soluble protein