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).External 2231691f894ba696de1310221b0a0dbbb31a7251e75115c265587c3d9d5f507c

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."]
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: 24036102External 2231691f894ba696de1310221b0a0dbbb31a7251e75115c265587c3d9d5f507c
Screening Toggle 893349bafcc528f8346c51dc3420151d67b0126b2c122dd1017121c03fa0f69b
  Experimental screening Non-experimental screening Reference
TRP channel construct Interactor source
TRP channel Interactor Method Species Region Species Organ/tissue Sample type
TRPV1 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 Actin Affinity purification-mass spectrometry Not used Mouse Dorsal root ganglion tissue Lysates 24036102
TRPV1 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 Apolipoprotein O Affinity purification-mass spectrometry Not used Mouse Dorsal root ganglion tissue Lysates 24036102
TRPV1 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 E-cadherin Affinity purification-mass spectrometry Not used Mouse Dorsal root ganglion tissue Lysates 24036102
TRPV1 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 Glucose 6 phosphatase Affinity purification-mass spectrometry Not used Mouse Dorsal root ganglion tissue Lysates 24036102
TRPV1 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 Glycogen phosphorylase Affinity purification-mass spectrometry Not used Mouse Dorsal root ganglion tissue Lysates 24036102
TRPV1 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 KvƢ2 Affinity purification-mass spectrometry Not used Mouse Dorsal root ganglion tissue Lysates 24036102
TRPV1 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 ơII-spectrin Affinity purification-mass spectrometry Not used Mouse Dorsal root ganglion tissue Lysates 24036102
TRPV1 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 RAB8b Affinity purification-mass spectrometry Not used Mouse Dorsal root ganglion tissue Lysates 24036102
TRPV1 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 Ribosomal protein L9 Affinity purification-mass spectrometry Not used Mouse Dorsal root ganglion tissue Lysates 24036102
TRPV1 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 snRNP 70 Affinity purification-mass spectrometry Not used Mouse Dorsal root ganglion tissue Lysates 24036102
TRPV1 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 TRPV2 Affinity purification-mass spectrometry Not used Mouse Dorsal root ganglion tissue Lysates 24036102
TRPV1 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 ZO-1 Affinity purification-mass spectrometry Not used Mouse Dorsal root ganglion tissue Lysates 24036102
TRPV2 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 TRPV1 Affinity purification-mass spectrometry Not used Mouse Dorsal root ganglion tissue Lysates 24036102
(Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4: click the arrow icon to show interactions only between the corresponding TRP channel and the interactor)
Validation: In vivo validation Toggle 893349bafcc528f8346c51dc3420151d67b0126b2c122dd1017121c03fa0f69b
  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 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 KvƢ2 Co-immunoprecipitation HEK293 tsA 201 Mouse Full-length Not specified Not specified 24036102
TRPV1 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 KvƢ2 Co-immunofluorescence staining Mouse dorsal root ganglion neuron 24036102
TRPV1 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 KvƢ2 Co-immunofluorescence staining HEK293 tsA 201 Mouse Full-length Not specified Not specified 24036102
(Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4: click the arrow icon to show interactions only between the corresponding TRP channel and the interactor)
Functional consequence Toggle 893349bafcc528f8346c51dc3420151d67b0126b2c122dd1017121c03fa0f69b
TRP channel Interactor Method Post-translational modification Subcellular trafficking Activity Reference
TRPV1 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 KvƢ2 Cell surface biotinylation Increase in plasma membrane level 24036102
(Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4: click the arrow icon to show interactions only between the corresponding TRP channel and the interactor)
TRP / Interactor

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