4.1 Article

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

Journal

BMC MOLECULAR AND CELL BIOLOGY
Volume 23, Issue 1, Pages -

Publisher

BMC
DOI: 10.1186/s12860-022-00405-w

Keywords

NanoBRET; C; elegans; Receptor-ligand interaction; Enhanced bystander BRET

Categories

Funding

  1. Free State of Saxony, Ministry for Higher Education, Research and the Arts (SMWK) [100316655]
  2. Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) [CRC 1423, 421152132]
  3. Projekt DEAL
  4. [FOR 2149]
  5. [265992651]

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The study demonstrates the use of in vivo NanoBRET as a powerful tool to investigate protein interactions and localization in a physiological setting in the living organism C. elegans.
Background Protein-protein interactions form the basis of every organism and thus, investigating their dynamics, intracellular protein localization, trafficking and interactions of distinct proteins such as receptors and their ligand-binding are of general interest. Bioluminescence resonance energy transfer (BRET) is a powerful tool to investigate these aspects in vitro. Since in vitro approaches mostly neglect the more complex in vivo situation, we established BRET as an in vivo tool for studying protein interactions in the nematode C. elegans. Results We generated worms expressing NanoBRET sensors and elucidated the interaction of two ligand-G protein-coupled receptor (GPCR) pairs, the neuropeptide receptor NPR-11 and the Adhesion GPCR LAT-1. Furthermore, we adapted the enhanced bystander BRET technology to measure subcellular protein localization. Using this approach, we traced ligand-induced internalization of NPR-11 in vivo. Conclusions Our results indicate that in vivo NanoBRET is a tool to investigate specific protein interactions and localization in a physiological setting in real time in the living organism C. elegans.

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