Identification of voltage-gated K+ channel beta 2 (Kv尾2) subunit as a novel interaction partner of the pain transducer Transient Receptor Potential Vanilloid 1 channel (TRPV1)
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- 作者：Carlo Bavassano ; Letizia Marvaldi ; Michiel Langeslag ; Bettina Sarg ; Herbert Lindner ; Lars Klimaschewski ; Michaela Kress ; Antonio Ferrer-Montiel ; Hans-G眉nther Knaus
- 关键词：AKAP ; A kinase anchor protein ; ATP ; adenosine tris-phosphate ; BSA ; bovine serum albumin ; cDNA ; complementary deoxyribonucleic acid ; CHAPS ; 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate ; CHO ; Chinese hamster ovary ; CGRP ; calcitonin gene-related peptide ; CMC ; critical micelle concentration ; DAPI ; 4&prime ; 6-diamidino-2-phenylindole ; DNA ; deoxyribonucleic acid ; DRG ; dorsal root ganglia ; DTT ; di-tiothreitol ; EDTA ; ethylenediaminetetraacetic acid ; EGTA ; ethylene glycol tetraacetic acid ; G
- 刊名：Biochimica et Biophysica Acta (BBA)/Molecular Cell Research
- 出版年：December, 2013
- 年：2013
- 卷：1833
- 期：12
- 页码：3166-3175
- 全文大小：953 K
文摘
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 (Kv尾2), 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/Kv尾2 association. Biotinylation assays in the presence of Kv尾2 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 Kv尾 subunit with TRPV1 channels, and suggests that such interaction may play a role in TRPV1 channel trafficking to the plasma membrane.