title: AlphaScreen selectivity assay for β-catenin/B-cell lymphoma 9 inhibitors.
abstract: The aberrant formation of the β-catenin/B-cell lymphoma 9 (BCL9) protein-protein complex is the driving force for many diseases, including cancer. Crystallographic analyses demonstrate that the surface area in β-catenin for interacting with BCL9 is overlapped with that for the β-catenin/E-cadherin interaction. In this study, a robust AlphaScreen selectivity assay was developed to quantify inhibitor potency for the β-catenin/BCL9 interaction and selectivity for β-catenin/BCL9 over β-catenin/E-cadherin interactions. A pilot screen was performed to demonstrat the feasibility of this assay. This selectivity assay is highly sensitive and suitable for adaptation to high-throughput screening. The establishment of this assay lays the foundation for the discovery of selective inhibitors specific for β-catenin/BCL9 interactions.
authors.affiliation: Department of Chemistry, Center for Cell and Genome Science, University of Utah, Salt Lake City, UT 84112, USA.
authors.forename: Min
authors.initials: M
authors.lastname: Zhang
authors.affiliation: Department of Chemistry, Center for Cell and Genome Science, University of Utah, Salt Lake City, UT 84112, USA.
authors.forename: John A
authors.initials: JA
authors.lastname: Wisniewski
authors.affiliation: Department of Chemistry, Center for Cell and Genome Science, University of Utah, Salt Lake City, UT 84112, USA. Electronic address: markji@chem.utah.edu.
authors.forename: Haitao
authors.initials: H
authors.lastname: Ji
mesh_terms: D045784:Abietanes / Q000032:analysis; D045744:Cell Line, Tumor; D004797:Enzyme-Linked Immunosorbent Assay; D005454:Fluorescence Polarization; D006801:Humans; D047468:Immunoprecipitation; D007700:Kinetics; D018613:Microscopy, Confocal; D009363:Neoplasm Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D010936:Plant Extracts / Q000032:analysis; D011485:Protein Binding; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D020349:Surface Plasmon Resonance; D014157:Transcription Factors; D051176:beta Catenin / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*
chemical_list: D045784:Abietanes; C111180:BCL9 protein, human; D009363:Neoplasm Proteins; D010936:Plant Extracts; D011994:Recombinant Proteins; D014157:Transcription Factors; D051176:beta Catenin; C018381:salvin
keywords: AlphaScreen; B-cell lymphoma 9; E-cadherin; Protein–protein interactions; Selectivity; β-Catenin