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Investigating Protein-protein Interactions in Live Cells Using Bioluminescence Resonance Energy Transfer
Published on: May 26, 2014
23.5K
Multimeric Purinoceptor Detection by Bioluminescence Resonance Energy Transfer
Vincent Compan1,2, François Rassendren3,4
1IGF, University of Montpellier, CNRS, INSERM, Montpellier, France. vincent.compan@igf.cnrs.fr.
Methods in Molecular Biology (Clifton, N.J.)
|October 25, 2019
Summary
Bioluminescence resonance energy transfer (BRET) assays offer a sensitive method to study protein interactions in living cells. This protocol details using BRET and bimolecular fluorescence complementation (BiFC) to investigate P2X subunit interactions and receptor stoichiometry.
Area of Science:
- Biochemistry
- Molecular Biology
- Cell Biology
Background:
- Protein-protein interactions are crucial for cellular function.
- Bioluminescence resonance energy transfer (BRET) is a sensitive technique for studying these interactions in real-time within live cells.
Purpose of the Study:
- To establish a protocol utilizing BRET to investigate potential interactions between P2X subunits.
- To explore the combined use of BRET and bimolecular fluorescence complementation (BiFC) for determining the stoichiometry of heteromeric P2X receptors.
Main Methods:
- Development and application of a BRET-based assay for live-cell analysis.
- Integration of BRET with bimolecular fluorescence complementation (BiFC) for stoichiometric analysis.
Main Results:
- Demonstration of a reliable BRET protocol for assessing P2X subunit interactions.
- Successful application of the combined BRET-BiFC approach to determine receptor stoichiometry.
Conclusions:
- BRET provides a robust method for studying protein-protein interactions, specifically P2X subunit interactions.
- The combined BRET-BiFC technique is effective for elucidating the subunit composition of heteromeric P2X receptors.
