Dataset Information

High-Throughput Selectivity Assays for Small-Molecule Inhibitors of ?-Catenin/T-Cell Factor Protein-Protein Interactions.

ABSTRACT: Two homogeneous high-throughput assays, AlphaScreen and fluorescence polarization, were established to quantify inhibitor selectivity between different protein-protein complexes. As a first case study, they have been successfully applied to the key protein-protein interactions in the downstream sites of the canonical Wnt signaling pathway. The aberrant formation of the ?-catenin/T-cell factor (Tcf) complex is the major driving force for many cancers and fibroses. Crystallographic and biochemical studies reveal that the binding modes of Tcf, E-cadherin, and adenomatous polyposis coli (APC) to ?-catenin are identical and mutually exclusive. In the present study, two highly sensitive and robust assays were established to quantitatively evaluate inhibitor selectivity between ?-catenin/Tcf, ?-catenin/E-cadherin, and ?-catenin/APC interactions. A pilot screen demonstrated the feasibility of the assays and yielded four hits for the disruption of ?-catenin/Tcf interactions. A potent and dual-selective ?-catenin/Tcf inhibitor was identified.

SUBMITTER: Zhang M

PROVIDER: S-EPMC4027448 | biostudies-literature | 2013 Feb

REPOSITORIES: biostudies-literature

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Publications

High-Throughput Selectivity Assays for Small-Molecule Inhibitors of β-Catenin/T-Cell Factor Protein-Protein Interactions.

Zhang Min M Catrow J Leon JL Ji Haitao H

ACS medicinal chemistry letters 20130108 2

Two homogeneous high-throughput assays, AlphaScreen and fluorescence polarization, were established to quantify inhibitor selectivity between different protein-protein complexes. As a first case study, they have been successfully applied to the key protein-protein interactions in the downstream sites of the canonical Wnt signaling pathway. The aberrant formation of the β-catenin/T-cell factor (Tcf) complex is the major driving force for many cancers and fibroses. Crystallographic and biochemical ...[more]

PMID: 24900664

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Project description:Protein-protein interactions play important roles in virtually all cellular processes, making them enticing targets for modulation by small-molecule therapeutics: specific examples have been well validated in diseases ranging from cancer and autoimmune disorders, to bacterial and viral infections. Despite several notable successes, however, overall these remain a very challenging target class. Protein interaction sites are especially challenging for computational approaches, because the target protein surface often undergoes a conformational change to enable ligand binding: this confounds traditional approaches for virtual screening. Through previous studies, we demonstrated that biased "pocket optimization" simulations could be used to build collections of low-energy pocket-containing conformations, starting from an unbound protein structure. Here, we demonstrate that these pockets can further be used to identify ligands that complement the protein surface. To do so, we first build from a given pocket its "exemplar": a perfect, but nonphysical, pseudoligand that would optimally match the shape and chemical features of the pocket. In our previous studies, we used these exemplars to quantitatively compare protein surface pockets to one another. Here, we now introduce this exemplar as a template for pharmacophore-based screening of chemical libraries. Through a series of benchmark experiments, we demonstrate that this approach exhibits comparable performance as traditional docking methods for identifying known inhibitors acting at protein interaction sites. However, because this approach is predicated on ligand/exemplar overlays, and thus does not require explicit calculation of protein-ligand interactions, exemplar screening provides a tremendous speed advantage over docking: 6 million compounds can be screened in about 15 min on a single 16-core, dual-GPU computer. The extreme speed at which large compound libraries can be traversed easily enables screening against a "pocket-optimized" ensemble of protein conformations, which in turn facilitates identification of more diverse classes of active compounds for a given protein target.

Dataset Information

High-Throughput Selectivity Assays for Small-Molecule Inhibitors of ?-Catenin/T-Cell Factor Protein-Protein Interactions.

Publications

High-Throughput Selectivity Assays for Small-Molecule Inhibitors of β-Catenin/T-Cell Factor Protein-Protein Interactions.

Similar Datasets

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