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Related Concept Videos

Ligand Binding Sites02:40

Ligand Binding Sites

Proteins are dynamic macromolecules that carry out a wide variety of essential processes; however, the activities of most proteins depend on their interactions with other molecules or ions, known as ligands.
Protein-ligand interactions are quite specific; even though numerous potential ligands surround a cellular protein at any given time, only a particular ligand can bind to that protein. Moreover, a ligand binds only to a dedicated area on the surface of the protein, known as the...
Conserved Binding Sites01:49

Conserved Binding Sites

Many proteins’ biological role depends on their interactions with their ligands, small molecules that bind to specific locations on the protein known as ligand-binding sites. Ligand-binding sites are often conserved among homologous proteins as these sites are critical for protein function.
Binding sites are often located in large pockets, and if their location on a protein’s surface is unknown, it can be predicted using various approaches. The energetic method computationally analyses the...
The Equilibrium Binding Constant and Binding Strength02:18

The Equilibrium Binding Constant and Binding Strength

The equilibrium binding constant (Kb) quantifies the strength of a protein-ligand interaction. Kb can be calculated as follows when the reaction is at equilibrium:
Assembly of Signaling Complexes01:30

Assembly of Signaling Complexes

Multiprotein signaling complexes are formed in a dynamic process involving protein-protein interactions at the cytoplasmic domain of transmembrane receptors or enzymatic and non-enzymatic proteins associated with the receptor. These complexes ensure the activation and propagation of intracellular signals that regulate cell functions.
Interaction domains in cell signaling
Interaction domains recognize exposed features of their binding partners containing post-translationally modified sequences,...

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Related Experiment Video

Updated: May 29, 2026

BRET-based G Protein Biosensors for Measuring G Protein-Coupled Receptor Activity in Live Cells
09:21

BRET-based G Protein Biosensors for Measuring G Protein-Coupled Receptor Activity in Live Cells

Published on: November 7, 2025

Using BRET to detect ligand-specific conformational changes in preformed signalling complexes.

Nicolas Audet1, Graciela Piñeyro

  • 1Department of Pharmacology, Faculty of Medicine, University of Montreal, Montreal, QC, Canada.

Methods in Molecular Biology (Clifton, N.J.)
|August 27, 2011
PubMed
Summary

Bioluminescence energy transfer (BRET) assays reveal that heptahelical receptors (7TMRs) and G proteins form stable signaling complexes. These complexes undergo ligand-specific conformational changes during receptor activation, providing insights into cellular signaling pathways.

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Area of Science:

  • Biochemistry
  • Cell Biology
  • Molecular Pharmacology

Background:

  • Bioluminescence energy transfer (BRET) is a key technique for studying protein interactions.
  • Heptahelical receptors (7TMRs) and heterotrimeric G proteins are crucial in cellular signaling.
  • Understanding signaling complex dynamics is vital for drug discovery.

Purpose of the Study:

  • To present protocols for BRET assays to study protein-protein interactions.
  • To demonstrate the existence of constitutive signaling complexes involving 7TMRs and proximal partners.
  • To investigate ligand-specific conformational changes within these complexes.

Main Methods:

  • Utilizing Bioluminescence energy transfer (BRET) assays in living cells.
  • Developing and applying protocols for BRET measurements.
  • Analyzing conformational changes in receptor-G protein signaling complexes.

Main Results:

  • BRET assays confirm the formation of constitutive signaling complexes by 7TMRs, G proteins, and effectors.
  • These multimeric protein arrays remain intact during initial receptor signaling stages.
  • Ligand binding induces distinct, ligand-specific intermolecular rearrangements in 7TMRs and G protein subunits.

Conclusions:

  • BRET is a powerful tool for dissecting the structure and dynamics of signaling complexes.
  • Constitutive signaling arrays provide a platform for direct information transfer upon receptor activation.
  • Ligand-specific conformational changes are central to the mechanism of 7TMR signaling.