NanoBRET in C. elegans illuminates functional receptor interactions in real time
Abstract 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 resonanc...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2022-01-01
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| Series: | BMC Molecular and Cell Biology |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12860-022-00405-w |
| _version_ | 1798023814125191168 |
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| author | Victoria Elisabeth Groß Miron Mikhailowitsch Gershkovich Torsten Schöneberg Anette Kaiser Simone Prömel |
| author_facet | Victoria Elisabeth Groß Miron Mikhailowitsch Gershkovich Torsten Schöneberg Anette Kaiser Simone Prömel |
| author_sort | Victoria Elisabeth Groß |
| collection | DOAJ |
| description | Abstract 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. |
| first_indexed | 2024-04-11T17:52:30Z |
| format | Article |
| id | doaj.art-c9f0a0db2abc4094be440a913a0ca113 |
| institution | Directory Open Access Journal |
| issn | 2661-8850 |
| language | English |
| last_indexed | 2024-04-11T17:52:30Z |
| publishDate | 2022-01-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Molecular and Cell Biology |
| spelling | doaj.art-c9f0a0db2abc4094be440a913a0ca1132022-12-22T04:11:01ZengBMCBMC Molecular and Cell Biology2661-88502022-01-0123111910.1186/s12860-022-00405-wNanoBRET in C. elegans illuminates functional receptor interactions in real timeVictoria Elisabeth Groß0Miron Mikhailowitsch Gershkovich1Torsten Schöneberg2Anette Kaiser3Simone Prömel4Rudolf Schönheimer Institute of Biochemistry, Medical Faculty, Leipzig UniversityInstitute of Biochemistry, Faculty of Life Sciences, Leipzig UniversityRudolf Schönheimer Institute of Biochemistry, Medical Faculty, Leipzig UniversityInstitute of Biochemistry, Faculty of Life Sciences, Leipzig UniversityRudolf Schönheimer Institute of Biochemistry, Medical Faculty, Leipzig UniversityAbstract 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.https://doi.org/10.1186/s12860-022-00405-wNanoBRETC. elegansReceptor-ligand interactionEnhanced bystander BRET |
| spellingShingle | Victoria Elisabeth Groß Miron Mikhailowitsch Gershkovich Torsten Schöneberg Anette Kaiser Simone Prömel NanoBRET in C. elegans illuminates functional receptor interactions in real time BMC Molecular and Cell Biology NanoBRET C. elegans Receptor-ligand interaction Enhanced bystander BRET |
| title | NanoBRET in C. elegans illuminates functional receptor interactions in real time |
| title_full | NanoBRET in C. elegans illuminates functional receptor interactions in real time |
| title_fullStr | NanoBRET in C. elegans illuminates functional receptor interactions in real time |
| title_full_unstemmed | NanoBRET in C. elegans illuminates functional receptor interactions in real time |
| title_short | NanoBRET in C. elegans illuminates functional receptor interactions in real time |
| title_sort | nanobret in c elegans illuminates functional receptor interactions in real time |
| topic | NanoBRET C. elegans Receptor-ligand interaction Enhanced bystander BRET |
| url | https://doi.org/10.1186/s12860-022-00405-w |
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