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Chemical library screening using a SPR-based inhibition in solution assay: simulations and experimental validation
Laurence Choulier1, Yves Nominé, Gabrielle Zeder-Lutz
1Biotechnologie et Signalisation Cellulaire, Université de Strasbourg , CNRS, ESBS, Boulevard Sébastien Brant BP10413, 67412 Illkirch, France.
Analytical Chemistry
|August 13, 2013
Summary
We developed a surface plasmon resonance assay to find inhibitors of protein interactions. This method effectively identified compounds that disrupt the binding between PDZ domains and peptides, suggesting optimal medium affinity for screening.
Area of Science:
- Biochemistry
- Molecular Biology
- Drug Discovery
Background:
- PDZ domains are crucial protein-protein interaction modules.
- Understanding interactions involving PDZ domains is key for therapeutic development.
- The E6 peptide and MAGI-1 PDZ1 interaction serves as a model system.
Purpose of the Study:
- To develop and validate a surface plasmon resonance (SPR)-based inhibition in solution assay (ISA).
- To screen a chemical library for inhibitors of the E6peptide-MAGI-1 PDZ1 interaction.
- To determine optimal conditions for SPR-based ISA screening.
Main Methods:
- Immobilization of E6-derived peptide on an SPR sensor.
- Utilizing a surface plasmon resonance (SPR) inhibition in solution assay (ISA).
- Screening the Prestwick Chemical Library® (1120 compounds) and performing concentration-dependent assays.
Main Results:
- Identified 36 compounds inhibiting the E6peptide-MAGI-1 PDZ1 interaction by over 5%.
- 13 compounds confirmed to affect the interaction in concentration-dependent ISA.
- Discovered three PDZ binders with dissociation constants (KD) in the 10-50 μM range.
Conclusions:
- SPR-based ISA is effective for identifying medium-affinity inhibitors.
- Medium (μM) affinity between target and surface-bound partner is optimal for SPR-based ISA.
- The developed assay facilitates the discovery of novel modulators of PDZ domain interactions.
