TRPV2 is a ligand-operated temperature sensor with poorly defined pharmacology. Here, we combine calcium imaging and patch-clamp electrophysiology with cryo-electron microscopy (cryo-EM) to explore how TRPV2 activity is modulated by the phytocannabinoid Δ9-tetrahydrocannabiorcol (C16) and by probenecid. C16 and probenecid act in concert to stimulate TRPV2 responses including histamine release from rat and human mast cells. Each ligand causes distinct conformational changes in TRPV2 as revealed by cryo-EM. Although the binding for probenecid remains elusive, C16 associates within the vanilloid pocket. As such, the C16 binding location is distinct from that of cannabidiol, partially overlapping with the binding site of the TRPV2 inhibitor piperlongumine. Taken together, we discover a new cannabinoid binding site in TRPV2 that is under the influence of allosteric control by probenecid. This molecular insight into ligand modulation enhances our understanding of TRPV2 in normal and pathophysiology.

Center for Biomolecular Magnetic Resonance Goethe University Frankfurt Main Germany

Department of Biomedical and Clinical Sciences Linköping University Linköping Sweden

Department of Biomedical Sciences University of Copenhagen Copenhagen Denmark

Department of Cellular Neurophysiology Institute of Physiology Czech Academy of Sciences Prague Czech Republic

Department of Clinical Sciences Malmö Lund University Malmö Sweden

Department of Experimental Medical Science Lund University Lund Sweden

Department of Molecular and Cellular Biology Harvard University Cambridge MA USA

Division Immunology and Allergy Unit Department of Medicine Solna Karolinska Institutet Karolinska University Hospital Solna Sweden

Faculty of Chemistry and Earth Sciences Institute of Organic Chemistry and Macromolecular Chemistry and Cluster of Excellence Balance of the Microverse Friedrich Schiller University Jena Jena Germany

Wallenberg Centre for Molecular Medicine Linköping University Linköping Sweden

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