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Protocol to assess receptor-ligand binding in C. elegans using adapted thermal shift assays
Samantha Y Tse1, Read Pukkila-Worley1
1Program in Innate Immunity, Division of Infectious Diseases and Immunology, Department of Medicine, UMass Chan Medical School, Worcester, MA 01655, USA.
STAR Protocols
|August 1, 2023
Summary
This study introduces a new protocol to test how well compounds bind to nuclear hormone receptors in Caenorhabditis elegans. This method helps identify potential drug targets by studying orphaned receptors.
Area of Science:
- Molecular biology
- Genomics
- Biochemistry
Background:
- The Caenorhabditis elegans genome has many nuclear hormone receptors, but many are not well understood.
- Identifying ligands for these orphaned receptors is crucial for understanding their function.
Purpose of the Study:
- To present a detailed protocol for assessing ligand-receptor binding in C. elegans.
- To enable the study of orphaned nuclear hormone receptors and their potential ligands.
Main Methods:
- Adapted cellular thermal shift assay (CETSA) and isothermal dose-response assays.
- Detailed steps for C. elegans cultivation, lysate preparation, and compound handling.
- Quantification methods for binding assays.
Main Results:
- The protocol provides a reproducible method for ligand-receptor binding assessment.
- Demonstrates applicability to soluble receptors in C. elegans.
Conclusions:
- This protocol facilitates the study of nuclear hormone receptor-ligand interactions in C. elegans.
- It offers a valuable tool for drug discovery and understanding basic biology.
