Related Experiment Video

Updated: Jun 27, 2026

Real-time Monitoring of Ligand-receptor Interactions with Fluorescence Resonance Energy Transfer
12:23

Real-time Monitoring of Ligand-receptor Interactions with Fluorescence Resonance Energy Transfer

Published on: August 20, 2012

Monitoring ligand-mediated nuclear receptor-coregulator interactions by noncovalent mass spectrometry

Sarah Sanglier1, William Bourguet, Pierre Germain

  • 1Laboratoire de Spectrométrie de Masse Bio-Organique, CNRS UMR 7509, ECPM, Strasbourg, France.

European Journal of Biochemistry
|December 21, 2004
PubMed

Insights

Nondenaturing electrospray ionization mass spectrometry (ESI-MS) reveals how different retinoids affect corepressor binding to retinoid receptors. This technique provides direct evidence for noncovalent nuclear receptor complexes and ligand binding characteristics.

Area of Science:

  • Molecular Biology
  • Biochemistry
  • Structural Biology

Background:

  • Nuclear receptors, including Retinoic Acid Receptors (RARs) and Retinoid X Receptors (RXRs), are crucial transcription factors.
  • These receptors mediate retinoid signaling by interacting with coactivators or corepressors at target gene promoters.
  • Corepressor interaction with nuclear receptors involves conserved LXXI/HIXXXI/L sequences known as CoRNR boxes.

Purpose of the Study:

  • To characterize CoRNR box peptide binding to the RARalpha-RXRalpha heterodimer using nondenaturing electrospray ionization mass spectrometry (ESI-MS).
  • To investigate the impact of various agonists, antagonists, and inverse agonists on the stability of RARalpha-RXRalpha-CoRNR ternary complexes.
  • To demonstrate the utility of ESI-MS in studying noncovalent nuclear receptor complexes and ligand-dependent interactions.

Main Methods:

  • Nondenaturing electrospray ionization mass spectrometry (ESI-MS) was employed to analyze peptide binding.
  • The stability of RARalpha-RXRalpha-CoRNR ternary complexes was monitored.
  • Different types of retinoid receptor ligands (agonists, antagonists, inverse agonists) were used to probe complex formation and stability.

Main Results:

  • ESI-MS provided direct evidence for the formation of noncovalent RARalpha-RXRalpha-CoRNR ternary complexes.
  • The study determined ligand binding stoichiometries within these complexes.
  • Differential effects of various retinoids on corepressor binding to the RARalpha-RXRalpha heterodimer were unambiguously shown.

Conclusions:

  • ESI-MS is a powerful technique for studying noncovalent nuclear receptor complexes, complementing traditional methods.
  • This mass spectrometry approach allows for direct observation and characterization of ligand-receptor-corepressor interactions.
  • The findings highlight the distinct influence of different retinoid ligands on the modulation of corepressor binding, offering insights into retinoid signaling pathways.

More Related Videos

A Protein Preparation Method for the High-throughput Identification of Proteins Interacting with a Nuclear Cofactor Using LC-MS/MS Analysis05:43

A Protein Preparation Method for the High-throughput Identification of Proteins Interacting with a Nuclear Cofactor Using LC-MS/MS Analysis

Published on: January 24, 2017

Measuring Interactions of Globular and Filamentous Proteins by Nuclear Magnetic Resonance Spectroscopy (NMR) and Microscale Thermophoresis (MST)10:28

Measuring Interactions of Globular and Filamentous Proteins by Nuclear Magnetic Resonance Spectroscopy (NMR) and Microscale Thermophoresis (MST)

Published on: November 2, 2018

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...
Ligand Binding and Linkage00:49

Ligand Binding and Linkage

Allosteric proteins have more than one ligand binding site; the binding of a ligand to any of these sites influences the binding of ligands to the other sites. When a protein is allosteric, its binding sites are called coupled or linked.  In the case of enzymes, the site that binds to the substrate is known as the active site and the other site is known as the regulatory site. When a ligand binds to the regulatory site, this leads to conformational changes in the protein that can influence the...
Ligand Binding and Linkage00:49

Ligand Binding and Linkage

Allosteric proteins have more than one ligand binding site; the binding of a ligand to any of these sites influences the binding of ligands to the other sites. When a protein is allosteric, its binding sites are called coupled or linked.  In the case of enzymes, the site that binds to the substrate is known as the active site and the other site is known as the regulatory site. When a ligand binds to the regulatory site, this leads to conformational changes in the protein that can influence the...
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...
Transducer Mechanism: Nuclear Receptors01:31

Transducer Mechanism: Nuclear Receptors

Nuclear receptors, or NRs, are unique transcription factors that regulate gene transcription and affect the cellular pathways involved in reproduction, development, or metabolism. Their ability to be stimulated by small lipophilic ligands and control vital cellular processes makes them ideal drug targets. Nearly 10-15% of currently prescribed drugs target these receptors.
About 48 different soluble family members of nuclear receptors are identified that can be divided into two main classes:
Protein-Drug Binding: Determination Methods01:22

Protein-Drug Binding: Determination Methods

Determining protein-drug binding can be achieved through indirect and direct methods, each providing valuable insights into the interaction between proteins and drugs.
Indirect methods involve isolating the bound drug from its free form in biological samples such as blood, serum, or plasma. These techniques aim to measure the percentage of drugs bound to proteins. Equilibrium dialysis is a commonly used method where the free drug concentration at equilibrium is measured by separating the bound...