Biochim Biophys Acta. 2013 Dec;1833(12):3166-75. doi: 10.1016/j.bbamcr.2013.09.001. Epub 2013 Sep 11.
Identification of voltage-gated K(+) channel beta 2 (Kvbeta2) subunit as a novel interaction partner of the pain transducer Transient Receptor Potential Vanilloid 1 channel (TRPV1).
Bavassano, C., Marvaldi, L., Langeslag, M., Sarg, B., Lindner, H., Klimaschewski, L., Kress, M., Ferrer-Montiel, A., Knaus, H. G.,
["Division of Cellular and Molecular Pharmacology, Medical University Innsbruck, Peter-Mayr strasse 1, 6020 Innsbruck, Austria. Electronic address: carlo.bavassano@i-med.ac.at."]
["Division of Cellular and Molecular Pharmacology, Medical University Innsbruck, Peter-Mayr strasse 1, 6020 Innsbruck, Austria. Electronic address: carlo.bavassano@i-med.ac.at."]
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 (Kvbeta2), 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/Kvbeta2 association. Biotinylation assays in the presence of Kvbeta2 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 Kvbeta subunit with TRPV1 channels, and suggests that such interaction may play a role in TRPV1 channel trafficking to the plasma membrane.
PMID: 24036102

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Screening
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Experimental screening | Non-experimental screening | Reference | ||||||||
TRP channel construct | Interactor source | |||||||||
TRP channel | Interactor | Method | Species | Region | Species | Organ/tissue | Sample type | |||
TRPV1 |
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Actin | Affinity purification-mass spectrometry | Not used | Mouse | Dorsal root ganglion tissue | Lysates | 24036102 | ||
TRPV1 |
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Apolipoprotein O | Affinity purification-mass spectrometry | Not used | Mouse | Dorsal root ganglion tissue | Lysates | 24036102 | ||
TRPV1 |
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E-cadherin | Affinity purification-mass spectrometry | Not used | Mouse | Dorsal root ganglion tissue | Lysates | 24036102 | ||
TRPV1 |
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Glucose 6 phosphatase | Affinity purification-mass spectrometry | Not used | Mouse | Dorsal root ganglion tissue | Lysates | 24036102 | ||
TRPV1 |
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Glycogen phosphorylase | Affinity purification-mass spectrometry | Not used | Mouse | Dorsal root ganglion tissue | Lysates | 24036102 | ||
TRPV1 |
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KvƢ2 | Affinity purification-mass spectrometry | Not used | Mouse | Dorsal root ganglion tissue | Lysates | 24036102 | ||
TRPV1 |
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ơII-spectrin | Affinity purification-mass spectrometry | Not used | Mouse | Dorsal root ganglion tissue | Lysates | 24036102 | ||
TRPV1 |
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RAB8b | Affinity purification-mass spectrometry | Not used | Mouse | Dorsal root ganglion tissue | Lysates | 24036102 | ||
TRPV1 |
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Ribosomal protein L9 | Affinity purification-mass spectrometry | Not used | Mouse | Dorsal root ganglion tissue | Lysates | 24036102 | ||
TRPV1 |
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snRNP 70 | Affinity purification-mass spectrometry | Not used | Mouse | Dorsal root ganglion tissue | Lysates | 24036102 | ||
TRPV1 |
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TRPV2 | Affinity purification-mass spectrometry | Not used | Mouse | Dorsal root ganglion tissue | Lysates | 24036102 | ||
TRPV1 |
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ZO-1 | Affinity purification-mass spectrometry | Not used | Mouse | Dorsal root ganglion tissue | Lysates | 24036102 | ||
TRPV2 |
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TRPV1 | Affinity purification-mass spectrometry | Not used | Mouse | Dorsal root ganglion tissue | Lysates | 24036102 |
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click the arrow icon to show interactions only between the corresponding TRP channel and the interactor)

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Validation: In vivo validation
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Assay with endogenous proteins | Assay with overexpressed proteins | Reference | ||||||||
Cell or tissue | Cell or tissue | TRP channel construct | Interactor construct | |||||||
TRP channel | Interactor | Method | Species | Region | Species | Region | ||||
TRPV1 |
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KvƢ2 | Co-immunoprecipitation | HEK293 tsA 201 | Mouse | Full-length | Not specified | Not specified | 24036102 | |
TRPV1 |
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KvƢ2 | Co-immunofluorescence staining | Mouse dorsal root ganglion neuron | 24036102 | |||||
TRPV1 |
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KvƢ2 | Co-immunofluorescence staining | HEK293 tsA 201 | Mouse | Full-length | Not specified | Not specified | 24036102 |
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click the arrow icon to show interactions only between the corresponding TRP channel and the interactor)

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Functional consequence
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TRP channel | Interactor | Method | Post-translational modification | Subcellular trafficking | Activity | Reference | ||||||
TRPV1 |
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KvƢ2 | Cell surface biotinylation | Increase in plasma membrane level | 24036102 |
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click the arrow icon to show interactions only between the corresponding TRP channel and the interactor)
