title: Targeting the transient receptor potential vanilloid type 1 (TRPV1) assembly domain attenuates inflammation-induced hypersensitivity.
abstract: The transient receptor potential channel vanilloid type 1 (TRPV1) is a non-selective cation channel expressed in sensory neurons of the dorsal root and trigeminal ganglia. TRPV1 is a polymodal channel activated by noxious heat, capsaicin, and protons. As a sensor for noxious stimuli, TRPV1 channel has been described as a key contributor to pain signaling. To form a functional channel, TRPV1 subunits must assemble into tetramers, and several studies have identified the TRPV1 C terminus as an essential element in subunit association. Here we combined biochemical assays with electrophysiology and imaging-based bimolecular fluorescence complementation (BiFC) and bioluminescence resonance energy transfer (BRET) in live cells to identify a short motif in the C-terminal tail of the TRPV1 subunit that governs channel assembly. Removing this region through early truncation or targeted deletion results in loss of subunit association and channel function. Importantly, we found that interfering with TRPV1 subunit association using a plasma membrane-tethered peptide attenuated mechanical and thermal hypersensitivity in two mouse models of inflammatory hyperalgesia. This represents a novel mechanism to disrupt TRPV1 subunit assembly and hence may offer a new analgesic tool for pain relief.
authors.affiliation: From the Department of Physiology and Pharmacology and Snyder Institute for Chronic Diseases, University of Calgary, Calgary, Alberta T2N 4N1, Canada and.
authors.forename: Robyn
authors.initials: R
authors.lastname: Flynn
authors.affiliation: From the Department of Physiology and Pharmacology and Snyder Institute for Chronic Diseases, University of Calgary, Calgary, Alberta T2N 4N1, Canada and.
authors.forename: Kevin
authors.initials: K
authors.lastname: Chapman
authors.affiliation: From the Department of Physiology and Pharmacology and Snyder Institute for Chronic Diseases, University of Calgary, Calgary, Alberta T2N 4N1, Canada and.
authors.forename: Mircea
authors.initials: M
authors.lastname: Iftinca
authors.affiliation: From the Department of Physiology and Pharmacology and Snyder Institute for Chronic Diseases, University of Calgary, Calgary, Alberta T2N 4N1, Canada and.
authors.forename: Reem
authors.initials: R
authors.lastname: Aboushousha
authors.affiliation: Centro de Estudios Moleculares de la Celula and Instituto de Ciencias Biomedicas, Facultad de Medicina, Universidad de Chile, Santiago, 8380453, Chile.
authors.forename: Diego
authors.initials: D
authors.lastname: Varela
authors.affiliation: From the Department of Physiology and Pharmacology and Snyder Institute for Chronic Diseases, University of Calgary, Calgary, Alberta T2N 4N1, Canada and altier@ucalgary.ca.
authors.forename: Christophe
authors.initials: C
authors.lastname: Altier
mesh_terms: D020816:Amino Acid Motifs; D000818:Animals; D001169:Arthritis, Experimental / Q000378:metabolism / Q000503:physiopathology; D001665:Binding Sites; D017353:Gene Deletion; D057809:HEK293 Cells; D006801:Humans; D006930:Hyperalgesia / Q000378:metabolism*; D007249:Inflammation / Q000378:metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D055503:Protein Multimerization; D021122:Protein Subunits / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D021381:Protein Transport; D051381:Rats; D050916:TRPV Cation Channels / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D021122:Protein Subunits; D050916:TRPV Cation Channels; C493922:Trpv1 protein, rat
keywords: Bioluminescence Resonance Energy Transfer (BRET); Biomolecular Fluorescence Complementation; Pain; Protein Assembly; Site-directed Mutagenesis; Trafficking; Transient Receptor Potential Channels (TRP Channels)