NanoBRET in C. elegans illuminates functional receptor interactions in real time

Area of Science:

• Molecular Biology
• Cell Biology
• Biophysics

Background:

• Protein-protein interactions are fundamental to all biological processes.
• Investigating protein dynamics, localization, and interactions is crucial for understanding cellular functions.
• Bioluminescence Resonance Energy Transfer (BRET) is effective for in vitro studies, but in vivo applications are less explored.

Purpose of the Study:

• To establish and validate NanoBRET as an in vivo tool for studying protein interactions and localization.
• To investigate ligand-GPCR interactions and receptor trafficking in a living organism.
• To adapt enhanced bystander BRET for real-time subcellular protein localization measurements.

Main Methods:

• Generation of Caenorhabditis elegans strains expressing NanoBRET sensors.
• Application of NanoBRET to study interactions between neuropeptide receptor NPR-11 and Adhesion GPCR LAT-1.
• Adaptation of enhanced bystander BRET technology for in vivo subcellular protein localization.

Main Results:

• Successfully elucidated interactions of neuropeptide receptor NPR-11 and Adhesion GPCR LAT-1 using in vivo NanoBRET.
• Demonstrated the capability of enhanced bystander BRET to measure subcellular protein localization in vivo.
• Traced ligand-induced internalization of NPR-11 in real time within C. elegans.

Conclusions:

• In vivo NanoBRET is a valuable tool for studying specific protein interactions in a physiological context.
• This method allows for real-time investigation of protein localization and dynamics in living organisms.
• The study highlights the potential of NanoBRET for advancing our understanding of biological processes in vivo.