title: Alpha-Based Multiplexed Assay for Identifying SH2 Domain Antagonists.
abstract: Constitutive activation of STAT3/5b frequently occurs in various human malignancies. STAT3/5b activation involves dimerization via intermolecular pTyr-SH2 binding; therefore, antagonizing this interaction is a feasible approach to inhibit STAT3/5b activation for cancer therapy. We have developed a multiplexed assay to assess STAT3- and STAT5b-SH2 binding in a single well by combining AlphaLISA and AlphaScreen beads. In this chapter, we describe application of the method for the purpose of identifying new STAT3 and STAT5b antagonists.
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 / Q000379:methods; D004305:Dose-Response Relationship, Drug; D055808:Drug Discovery / Q000379:methods; D057166:High-Throughput Screening Assays; D006801:Humans; D008024:Ligands; D011485:Protein Binding; D011506:Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry*; D050796:STAT3 Transcription Factor / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism; D050799:STAT5 Transcription Factor / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism; D054852:Small Molecule Libraries; D018909:src Homology Domains
chemical_list: D008024:Ligands; D011506:Proteins; D050796:STAT3 Transcription Factor; D050799:STAT5 Transcription Factor; D054852:Small Molecule Libraries
keywords: Alpha assay; High-throughput screening (HTS); Multiplexed assay; STAT3; STAT5b; Src homology 2 (SH2)