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