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Optical tools for visualizing and controlling human GLP-1 receptor activation with high spatiotemporal resolution


Duffet, Loïc; Williams, Elyse T; Gresch, Andrea; Chen, Simin; Bhat, Musadiq A; Benke, Dietmar; Hartrampf, Nina; Patriarchi, Tommaso (2023). Optical tools for visualizing and controlling human GLP-1 receptor activation with high spatiotemporal resolution. eLife, 12:RP86628.

Abstract

The glucagon-like peptide-1 receptor (GLP1R) is a broadly expressed target of peptide hormones with essential roles in energy and glucose homeostasis, as well as of the blockbuster weight-loss drugs semaglutide and liraglutide. Despite its large clinical relevance, tools to investigate the precise activation dynamics of this receptor with high spatiotemporal resolution are limited. Here, we introduce a novel genetically encoded sensor based on the engineering of a circularly permuted green fluorescent protein into the human GLP1R, named GLPLight1. We demonstrate that fluorescence signal from GLPLight1 accurately reports the expected receptor conformational activation in response to pharmacological ligands with high sensitivity (max ΔF/F$_{0}$=528%) and temporal resolution (τ$_{ON}$ = 4.7 s). We further demonstrated that GLPLight1 shows comparable responses to glucagon-like peptide-1 (GLP-1) derivatives as observed for the native receptor. Using GLPLight1, we established an all-optical assay to characterize a novel photocaged GLP-1 derivative (photo-GLP1) and to demonstrate optical control of GLP1R activation. Thus, the new all-optical toolkit introduced here enhances our ability to study GLP1R activation with high spatiotemporal resolution.

Abstract

The glucagon-like peptide-1 receptor (GLP1R) is a broadly expressed target of peptide hormones with essential roles in energy and glucose homeostasis, as well as of the blockbuster weight-loss drugs semaglutide and liraglutide. Despite its large clinical relevance, tools to investigate the precise activation dynamics of this receptor with high spatiotemporal resolution are limited. Here, we introduce a novel genetically encoded sensor based on the engineering of a circularly permuted green fluorescent protein into the human GLP1R, named GLPLight1. We demonstrate that fluorescence signal from GLPLight1 accurately reports the expected receptor conformational activation in response to pharmacological ligands with high sensitivity (max ΔF/F$_{0}$=528%) and temporal resolution (τ$_{ON}$ = 4.7 s). We further demonstrated that GLPLight1 shows comparable responses to glucagon-like peptide-1 (GLP-1) derivatives as observed for the native receptor. Using GLPLight1, we established an all-optical assay to characterize a novel photocaged GLP-1 derivative (photo-GLP1) and to demonstrate optical control of GLP1R activation. Thus, the new all-optical toolkit introduced here enhances our ability to study GLP1R activation with high spatiotemporal resolution.

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Additional indexing

Item Type:Journal Article, refereed, original work
Communities & Collections:04 Faculty of Medicine > Institute of Pharmacology and Toxicology
07 Faculty of Science > Institute of Pharmacology and Toxicology

07 Faculty of Science > Department of Chemistry
04 Faculty of Medicine > Neuroscience Center Zurich
Dewey Decimal Classification:540 Chemistry
610 Medicine & health
570 Life sciences; biology
Uncontrolled Keywords:General Immunology and Microbiology, General Biochemistry, Genetics and Molecular Biology, General Medicine, General Neuroscience
Language:English
Date:2 June 2023
Deposited On:18 Feb 2024 15:34
Last Modified:18 Apr 2024 03:50
Publisher:eLife Sciences Publications Ltd.
ISSN:2050-084X
OA Status:Gold
Free access at:Publisher DOI. An embargo period may apply.
Publisher DOI:https://doi.org/10.7554/elife.86628.3
Project Information:
  • : FunderH2020
  • : Grant ID891959
  • : Project TitleOPTONEUROCHEM - Genetically encoded sensors for imaging neurochemical dynamics in vivo
  • : FunderSNF
  • : Grant ID10030_196455
  • : Project Title
  • : FunderSNF
  • : Grant ID310030L_212508
  • : Project Title
  • : FunderSNF
  • : Grant ID200021_200865
  • : Project Title
  • Content: Published Version
  • Language: English
  • Licence: Creative Commons: Attribution 4.0 International (CC BY 4.0)