Novel Method for Quantifying AhR-Ligand Binding Affinities Using Microscale Thermophoresis.

Area of Science:

• Biochemistry and Molecular Biology
• Pharmacology and Drug Discovery

Background:

• The aryl hydrocarbon receptor (AhR) is a crucial cellular sensor for environmental pollutants and endogenous metabolites.
• AhR pathway dysregulation is linked to diseases like cancer and autoimmune disorders, making it a therapeutic target.
• Understanding AhR ligand binding is essential for developing targeted therapies and selective AhR modulators.

Purpose of the Study:

• To present a novel microscale thermophoresis-based assay for monitoring AhR ligand binding.
• To enable precise identification and characterization of unknown AhR ligands in a cell-free system.
• To establish a screening strategy for discovering selective AhR modulators.

Main Methods:

• Utilized microscale thermophoresis (MST) to measure binding interactions.
• Employed purified recombinant human AhR in a cell-free assay.
• Monitored the binding of AhR to its natural ligands.

Main Results:

• Successfully demonstrated a novel MST-based approach for AhR ligand binding analysis.
• The method allows for precise characterization of ligand interactions with purified AhR.
• Validated the approach for identifying and characterizing unknown AhR ligands.

Conclusions:

• The developed microscale thermophoresis assay is effective for studying AhR ligand binding.
• This method facilitates the discovery of novel AhR ligands and potential selective modulators.
• The approach supports targeted drug development for AhR-related diseases.