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New homogeneous high-throughput assays for inhibitors of β-catenin/Tcf protein-protein interactions
Min Zhang1, Zheng Huang, Binxun Yu
1Department of Chemistry, Center for Cell and Genome Science, University of Utah, Salt Lake City, UT 84112-0850, USA.
Abstract:
The transcriptional activator β-catenin is the primary mediator of the canonical Wnt/β-catenin signaling pathway. The aberrant formation of the β-catenin/T-cell factor (Tcf) complex leads to many cancers and organ fibroses. Selective inhibition of β-catenin/Tcf protein-protein interactions represents an appealing therapeutic strategy. In this study, two new robust, homogeneous high-throughput assays, AlphaScreen and fluorescence polarization, were established to study β-catenin/Tcf interactions. These two new assays reproduce native β-catenin/Tcf interactions, quantify inhibitory potency, and are complementary with each other. The establishment of these two high-throughput screenings enables quick identification of inhibitors of β-catenin/Tcf protein-protein interactions.
Insights
Researchers developed two new high-throughput assays to identify inhibitors of the beta-catenin/T-cell factor (Tcf) complex, crucial in cancer and fibrosis. These assays enable rapid screening for potential therapeutic strategies targeting these protein-protein interactions.
Area of Science:
- Molecular Biology
- Biochemistry
- Drug Discovery
Background:
- The canonical Wnt/β-catenin signaling pathway is regulated by the transcriptional activator β-catenin.
- Aberrant β-catenin/T-cell factor (Tcf) complex formation is implicated in various cancers and organ fibroses.
- Targeting β-catenin/Tcf protein-protein interactions offers a promising therapeutic avenue.
Purpose of the Study:
- To establish robust, homogeneous high-throughput assays for studying β-catenin/Tcf interactions.
- To enable the quantification of inhibitory potency against the β-catenin/Tcf complex.
- To facilitate the rapid identification of novel inhibitors for therapeutic development.
Main Methods:
- Development and validation of AlphaScreen assay.
- Development and validation of fluorescence polarization assay.
- Utilizing these assays to study β-catenin/Tcf protein-protein interactions and inhibitor screening.
Main Results:
- Two new high-throughput assays (AlphaScreen and fluorescence polarization) were successfully established.
- These assays accurately reproduce native β-catenin/Tcf interactions.
- The assays demonstrated complementary capabilities in quantifying inhibitory potency.
Conclusions:
- The developed high-throughput assays are effective tools for studying β-catenin/Tcf interactions.
- These assays facilitate the rapid screening and identification of potential inhibitors.
- This work supports the development of new therapeutic strategies for diseases driven by aberrant Wnt/β-catenin signaling.
