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

10120 Department of Chemistry 0301 basic medicine QH301-705.5 Science 11558 Neuroscience Center Zurich 10050 Institute of Pharmacology and Toxicology 610 Medicine & health Genetics and Molecular Biology Glucagon-Like Peptide-1 Receptor 03 medical and health sciences Biochemistry and Chemical Biology Glucagon-Like Peptide 1 540 Chemistry Humans GLP1R Biology (General) neuropeptide genetically encoded sensor General Immunology and Microbiology General Neuroscience Q R General Medicine photocage Liraglutide General Biochemistry 570 Life sciences; biology Medicine fluorescence
DOI: 10.7554/elife.86628 Publication Date: 2023-04-11T10:49:37Z
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/F0=528%) and temporal resolution (τON = 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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