biostudies-literatureLiang DCharlotte Research Institute, University of North Carolina at CharlotteNIDDK NIH HHSAlfred P Sloan FoundationNational Institutes of HealthCancer Research InstituteCornell UniversityNational Science Foundation Division of ChemistryArnold and Mabel Beckman FoundationChulabhorn Research InstituteNational Science Foundation67-73.e3https://www.ebi.ac.uk/biostudies/studies/S-EPMC8655016biostudies-literatureUnknown29(1)Canonically, G-protein-coupled receptor (GPCR) signaling is transient and confined to the plasma membrane (PM). Deviating from this paradigm, the parathyroid hormone receptor (PTHR1) stimulates sustained G_s signaling at endosomes. In addition to G_s, PTHR1 activates G_q signaling; yet, in contrast to the PTHR1-G_s pathway, the spatiotemporal dynamics of the G_q branch of PTHR1 signaling and its relationship to G_s signaling remain largely ill defined. Recognizing that a downstream consequence of G_q signaling is the activation of phospholipase D (PLD) enzymes, we leverage activity-based, bioorthogonal imaging tools for PLD signaling to visualize and quantify the G_q branch of PTHR1 signaling. We establish thatCell chemical biologyActivity-based, bioorthogonal imaging of phospholipase D reveals spatiotemporal dynamics of GPCR-Gq signaling.PMC8655016P01DK011794CHE-1749919P01 DK011794Liang DGardella TJWatanabe TCheloha RWBaskin JMfalseActivity-based, bioorthogonal imaging of phospholipase D reveals spatiotemporal dynamics of GPCR-Gq signaling.Canonically, G-protein-coupled receptor (GPCR) signaling is transient and confined to the plasma membrane (PM). Deviating from this paradigm, the parathyroid hormone receptor (PTHR1) stimulates sustained G_s signaling at endosomes. In addition to G_s, PTHR1 activates G_q signaling; yet, in contrast to the PTHR1-G_s pathway, the spatiotemporal dynamics of the G_q branch of PTHR1 signaling and its relationship to G_s signaling remain largely ill defined. Recognizing that a downstream consequence of G_q signaling is the activation of phospholipase D (PLD) enzymes, we leverage activity-based, bioorthogonal imaging tools for PLD signaling to visualize and quantify the G_q branch of PTHR1 signaling. We establish that2022-01-01T00:00:00Z2022 Jan2025-04-04T13:40:38.791Z2024-11-09T02:23:06.714ZS-EPMC86550163416178610.1016/j.chembiol.2021.05.020