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Reverse MAPPIT: screening for protein-protein interaction modifiers in mammalian cells.
Sven Eyckerman1, Irma Lemmens, Dominiek Catteeuw
1Flanders Interuniversity Institute for Biotechnology, VIB09, Department of Medical Protein Research, Faculty of Medicine and Health Sciences, Ghent University, A. Baertsoenkaai 3, B-9000 Ghent, Belgium.
Nature Methods
|May 24, 2005
Summary
This study introduces a novel assay to identify molecules disrupting protein-protein interactions crucial for cellular function and disease. The reverse mammalian protein-protein interaction trap (MAPPIT) assay enables screening for potential therapeutics targeting these interactions.
Area of Science:
- Biochemistry
- Cell Biology
- Pharmacology
Background:
- Protein-protein interactions (PPIs) are fundamental to cellular processes.
- Dysregulated PPIs are implicated in various diseases and pathogen mechanisms.
- Targeting PPIs offers therapeutic potential but requires effective screening tools.
Purpose of the Study:
- To develop and evaluate a novel cytokine receptor-based assay for screening PPI disruptors.
- To demonstrate the utility of the reverse mammalian protein-protein interaction trap (MAPPIT) assay in intact mammalian cells.
- To validate the MAPPIT assay using known protein interaction pairs.
Main Methods:
- Development of a positive readout assay based on cytokine receptor signaling.
- Implementation of the reverse mammalian protein-protein interaction trap (MAPPIT) system.
- Screening of polypeptide and small organic molecules as PPI disruptors.
- Validation using specific PPIs: EpoR-SOCS, FKBP12-ALK4, and MDM2-p53.
Main Results:
- Successfully established and validated the MAPPIT assay for monitoring PPIs in live cells.
- Demonstrated the assay's capability to detect disruptors of specific PPIs.
- Evaluated the assay's performance with both protein fragments and small molecules.
Conclusions:
- The MAPPIT assay provides a robust platform for identifying inhibitors of protein-protein interactions.
- This assay facilitates drug discovery efforts targeting PPIs in various diseases.
- The MAPPIT system is adaptable for studying diverse protein interactions within mammalian cells.