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Ligand-based receptor identification on living cells and tissues using TRICEPS
Andreas P Frei1, Hansjoerg Moest, Karel Novy
1Department of Biology, Institute of Molecular Systems Biology, ETH Zurich, Zurich, Switzerland.
Nature Protocols
|June 15, 2013
Summary
TRICEPS is a novel chemoproteomic reagent enabling ligand-based receptor capture (LRC) on living cells. This method sensitively identifies specific receptors for various ligands using quantitative mass spectrometry.
Area of Science:
- Chemical Biology
- Proteomics
- Cell Biology
Background:
- Cell surface protein receptors mediate physiological responses to diverse ligands.
- Identifying specific ligand-receptor interactions is crucial for understanding cellular signaling and drug development.
Purpose of the Study:
- To introduce TRICEPS, a chemoproteomic reagent for ligand-based receptor capture (LRC).
- To enable unbiased and sensitive identification of specific receptors for ligands on living cells and tissues.
Main Methods:
- TRICEPS couples to ligands, capturing glycosylated receptors on oxidized cells via orthogonal functionalities.
- Biotinylation and quantitative mass spectrometry (MS) are used for receptor peptide purification and analysis.
- Comparative analysis with control probes identifies specific ligand-receptor interactions.
Main Results:
- TRICEPS facilitates sensitive, unbiased identification of specific receptors for various ligands.
- The method works under near-physiological conditions without genetic manipulation.
- TRICEPS can identify approximate ligand-binding sites for small ligands.
Conclusions:
- Ligand-based receptor capture (LRC) using TRICEPS is a powerful technology for receptor identification.
- This approach offers a rapid (within 1 week) and versatile tool for biological research and drug discovery.
