title: Identification of voltage-gated K(+) channel beta 2 (Kvβ2) subunit as a novel interaction partner of the pain transducer Transient Receptor Potential Vanilloid 1 channel (TRPV1).
abstract: The Transient Receptor Potential Vanilloid 1 (TRPV1, vanilloid receptor 1) ion channel plays a key role in the perception of thermal and inflammatory pain, however, its molecular environment in dorsal root ganglia (DRG) is largely unexplored. Utilizing a panel of sequence-directed antibodies against TRPV1 protein and mouse DRG membranes, the channel complex from mouse DRG was detergent-solubilized, isolated by immunoprecipitation and subsequently analyzed by mass spectrometry. A number of potential TRPV1 interaction partners were identified, among them cytoskeletal proteins, signal transduction molecules, and established ion channel subunits. Based on stringent specificity criteria, the voltage-gated K(+) channel beta 2 subunit (Kvβ2), an accessory subunit of voltage-gated K(+) channels, was identified of being associated with native TRPV1 channels. Reverse co-immunoprecipitation and antibody co-staining experiments confirmed TRPV1/Kvβ2 association. Biotinylation assays in the presence of Kvβ2 demonstrated increased cell surface expression levels of TRPV1, while patch-clamp experiments resulted in a significant increase of TRPV1 sensitivity to capsaicin. Our work shows, for the first time, the association of a Kvβ subunit with TRPV1 channels, and suggests that such interaction may play a role in TRPV1 channel trafficking to the plasma membrane.
authors.affiliation: Division of Cellular and Molecular Pharmacology, Medical University Innsbruck, Peter-Mayr strasse 1, 6020 Innsbruck, Austria. Electronic address: carlo.bavassano@i-med.ac.at.
authors.forename: Carlo
authors.initials: C
authors.lastname: Bavassano
authors.affiliation: Division of Neuroanatomy, Medical University Innsbruck, Müllerstrasse 59, 6020 Innsbruck, Austria. Electronic address: letizia.marvaldi@i-med.ac.at.
authors.forename: Letizia
authors.initials: L
authors.lastname: Marvaldi
authors.affiliation: Division of Physiology, Medical University Innsbruck, Fritz-Pregl-Straße 3, 6020 Innsbruck, Austria. Electronic address: michiel.langeslag@i-med.ac.at.
authors.forename: Michiel
authors.initials: M
authors.lastname: Langeslag
authors.affiliation: Division of Clinical Biochemistry, Medical University Innsbruck, Innrain 80, 6020 Innsbruck, Austria. Electronic address: bettina.sarg@i-med.ac.at.
authors.forename: Bettina
authors.initials: B
authors.lastname: Sarg
authors.affiliation: Division of Clinical Biochemistry, Medical University Innsbruck, Innrain 80, 6020 Innsbruck, Austria. Electronic address: herbert.lindner@i-med.ac.at.
authors.forename: Herbert
authors.initials: H
authors.lastname: Lindner
authors.affiliation: Division of Neuroanatomy, Medical University Innsbruck, Müllerstrasse 59, 6020 Innsbruck, Austria. Electronic address: lars.klimaschewski@i-med.ac.at.
authors.forename: Lars
authors.initials: L
authors.lastname: Klimaschewski
authors.affiliation: Division of Physiology, Medical University Innsbruck, Fritz-Pregl-Straße 3, 6020 Innsbruck, Austria. Electronic address: michaela.kress@i-med.ac.at.
authors.forename: Michaela
authors.initials: M
authors.lastname: Kress
authors.affiliation: IBMC, Universidad Miguel Hernandez Elche, Av. de la Universidad s/n., Edif. Torregaitán, E-03202, Spain. Electronic address: aferrer@umh.es.
authors.forename: Antonio
authors.initials: A
authors.lastname: Ferrer-Montiel
authors.affiliation: Division of Cellular and Molecular Pharmacology, Medical University Innsbruck, Peter-Mayr strasse 1, 6020 Innsbruck, Austria. Electronic address: hans.g.knaus@i-med.ac.at.
authors.forename: Hans-Günther
authors.initials: HG
authors.lastname: Knaus
mesh_terms: D000818:Animals; D019921:Biotinylation; D002462:Cell Membrane / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D047468:Immunoprecipitation; D013058:Mass Spectrometry; D051379:Mice; D018345:Mice, Knockout; D018408:Patch-Clamp Techniques; D011485:Protein Binding; D021122:Protein Subunits / Q000378:metabolism*; D051381:Rats; D011994:Recombinant Proteins / Q000378:metabolism; D051637:Shaker Superfamily of Potassium Channels / Q000737:chemistry / Q000378:metabolism*; D050916:TRPV Cation Channels / Q000378:metabolism*
chemical_list: C101328:Kcnab2 protein, mouse; D021122:Protein Subunits; D011994:Recombinant Proteins; D051637:Shaker Superfamily of Potassium Channels; D050916:TRPV Cation Channels; C493921:TRPV1 protein, mouse
keywords: 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate; 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid; 4′,6-diamidino-2-phenylindole; A kinase anchor protein; AKAP; ATP; Accessory subunit; BSA; CGRP; CHAPS; CHO; CMC; Chinese hamster ovary; DAPI; DNA; DRG; DTT; Dorsal root ganglia; EDTA; EGTA; GAPDH; HEPES; Icaps; IgG; Kvβ2; PBS; PMSF; SDS; Signaling complex; TRPV1; Transient Receptor Potential Vanilloid 1; WT mouse; adenosine tris-phosphate; bovine serum albumin; cDNA; calcitonin gene-related peptide; capsaicin-activated current; complementary deoxyribonucleic acid; critical micelle concentration; deoxyribonucleic acid; di-tiothreitol; dorsal root ganglia; ethylene glycol tetraacetic acid; ethylenediaminetetraacetic acid; glyceraldehyde-3-phosphate dehydrogenase; immunoglobulin G; phenylmethylsulfonyl fluoride; phosphate buffer saline; sodium dodecyl sulfate; voltage-gated K(+) channel subunit beta 2; wild type mouse