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A yeast sensor of ligand binding.

C L Tucker¹, S Fields

¹Departments of Genetics and Medicine, University of Washington, Box 357360, Seattle, WA 98195, USA.

Nature Biotechnology

|November 2, 2001

Summary

This study introduces a novel yeast biosensor for detecting small-molecule protein interactions. The system measures changes in yeast growth to identify new ligands and binding domains, offering a simple and cost-effective screening method.

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Area of Science:

Biotechnology
Molecular Biology
Synthetic Biology

Background:

Developing sensitive and efficient biosensors is crucial for drug discovery and understanding protein-ligand interactions.
Existing methods for ligand screening can be complex and costly.

Purpose of the Study:

To develop a novel yeast-based biosensor for detecting small-molecule ligand binding to proteins.
To demonstrate the utility of this biosensor for identifying novel ligands, ligand-binding domains, and mutations affecting protein activity.

Main Methods:

Engineered temperature-sensitive yeast strains lacking dihydrofolate reductase (DHFR).
Complemented DHFR activity using mouse DHFR fused to ligand-binding domains (FKBP12 and estrogen receptor-alpha [ERalpha]).
Measured changes in yeast growth in response to ligand binding.

Main Results:

Yeast strains expressing FKBP12-DHFR and ERalpha-DHFR fusions showed ligand-dependent growth.
The biosensor identified mutations affecting ERalpha ligand binding and protein activity.
Screening against a chemical array identified ligands binding to FKBP12 and ERalpha.
The ERalpha sensor differentiated between estrogen analogs based on their binding affinities.

Conclusions:

The developed yeast growth assay is a simple, inexpensive, and effective method for screening small-molecule ligands and identifying novel ligand-binding domains.
This biosensor platform has broad applications in drug discovery and chemical biology.

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