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Updated: May 11, 2026
Real Time Monitoring of Intracellular Bile Acid Dynamics Using a Genetically Encoded FRET-based Bile Acid Sensor
Published on: January 4, 2016
Development of time resolved fluorescence resonance energy transfer-based assay for FXR antagonist discovery
Donna D Yu1, Wenwei Lin, Taosheng Chen
1Department of Diabetes, Endocrinology and Metabolism, Beckman Research Institute, City of Hope National Medical Center, Duarte, CA 91010, USA. dyu@coh.org
Abstract:
FXR (farnesoid X receptor, NRIH4), a nuclear receptor, plays a major role in the control of cholesterol metabolism. FXR ligands have been investigated in preclinical studies for targeted therapy against metabolic diseases, but have shown limitations. Therefore, there is a need for new agonist or antagonist ligands of FXR, both for potential clinical applications, as well as to further elucidate its biological functions. Here we describe the use of the X-ray crystal structure of FXR complexed with the potent small molecule agonist GW4064 to design and synthesize a novel fluorescent, high-affinity probe (DY246) for time resolved fluorescence resonance energy transfer (TR-FRET) assays. We then used the TR-FRET assay for high throughput screening of a library of over 5000 bioactive compounds. From this library, we identified 13 compounds that act as putative FXR transcriptional antagonists.
Insights
Researchers developed a novel fluorescent probe to screen over 5000 compounds for farnesoid X receptor (FXR) antagonists. This led to the identification of 13 potential FXR transcriptional antagonists for metabolic disease research.
Area of Science:
- Biochemistry
- Molecular Biology
- Pharmacology
Background:
- The farnesoid X receptor (FXR) is a nuclear receptor crucial for regulating cholesterol metabolism.
- Current FXR ligands show limitations in preclinical studies for metabolic diseases, necessitating new therapeutic agents.
- Understanding FXR's biological functions requires novel tools and ligands.
Purpose of the Study:
- To design and synthesize a high-affinity fluorescent probe for FXR.
- To develop a time-resolved fluorescence resonance energy transfer (TR-FRET) assay for FXR.
- To perform high-throughput screening (HTS) to identify novel FXR antagonists.
Main Methods:
- Utilized the X-ray crystal structure of FXR complexed with the agonist GW4064 for probe design.
- Synthesized a novel fluorescent probe (DY246) for TR-FRET assays.
- Screened a library of over 5000 bioactive compounds using the developed TR-FRET assay.
Main Results:
- Successfully designed and synthesized a high-affinity fluorescent probe, DY246.
- Established a robust TR-FRET assay for FXR activity.
- Identified 13 compounds exhibiting putative FXR transcriptional antagonist activity from the screened library.
Conclusions:
- The novel fluorescent probe and TR-FRET assay are effective tools for FXR research.
- The identified compounds represent potential leads for developing new therapies targeting metabolic diseases via FXR antagonism.
- Further investigation of these 13 compounds is warranted to validate their FXR antagonist potential and therapeutic utility.
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