J Biol Chem. 2004 Jun 11;279(24):25665-72. Epub 2004 Apr 5.
Regulated exocytosis contributes to protein kinase C potentiation of vanilloid receptor activity.
Morenilla-Palao, C., Planells-Cases, R., Garcia-Sanz, N., Ferrer-Montiel, A.,
["Instituto de Biologia Molecular y Celular, Universidad Miguel Hernandez, Avenida del Ferrocarril s/n, 03202 Elche (Alicante), Spain."]
["Instituto de Biologia Molecular y Celular, Universidad Miguel Hernandez, Avenida del Ferrocarril s/n, 03202 Elche (Alicante), Spain."]
The vanilloid receptor-1 (TRPV1) plays a key role in the perception of peripheral thermal and inflammatory pain. TRPV1 expression and channel activity are notably up-regulated by proalgesic agents. The transduction pathways involved in TRPV1 sensitization are still elusive. We have used a yeast two-hybrid screen to identify proteins that associate with the N terminus of TRPV1. We report that two vesicular proteins, Snapin and synaptotagmin IX (Syt IX), strongly interact in vitro and in vivo with the TRPV1 N-terminal domain. In primary dorsal root ganglion neurons, TRPV1 co-distributes in vesicles with Syt IX and the vesicular protein synaptobrevin. Neither Snapin nor Syt IX affected channel function, but they notably inhibited protein kinase C (PKC)-induced potentiation of TRPV1 channel activity with a potency that rivaled the blockade evoked by botulinum neurotoxin A, a potent blocker of neuronal exocytosis. Noteworthily, we found that PKC activation induced a rapid delivery of functional TRPV1 channels to the plasma membrane. Botulinum neurotoxin A blocked the TRPV1 membrane translocation induced by PKC that was activated with a phorbol ester or the metabotropic glutamate receptor mGluR5. Therefore, our results indicate that PKC signaling promotes at least in part the SNARE-dependent exocytosis of TRPV1 to the cell surface. Taken together, these findings imply that activity-dependent delivery of channels to the neuronal surface may contribute to the buildup and maintenance of thermal inflammatory hyperalgesia in peripheral nociceptor terminals.
PMID: 15066994
 Screening
Validation: In vitro validation
Validation: In vivo validation
Characterization
Functional consequence
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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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SNAPIN | Yeast two-hybrid | Rat | 1-414 | Rat | Brain | cDNA library | 15066994 | |
| TRPV1 |
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Synaptotagmin 1 | Inference | Prediction | 15066994 | |||||
| TRPV1 |
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Synaptotagmin 4 | Yeast two-hybrid | Rat | 1-414 | Rat | Brain | cDNA library | 15066994 | |
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click the arrow icon to show interactions only between the corresponding TRP channel and the interactor)
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click the arrow icon to show interactions only between the corresponding TRP channel and the interactor)
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Screening
Validation: In vitro validation
Validation: In vivo validation
Characterization
Functional consequence
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top
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| Validation: In vitro validation
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| Assay with recombinant proteins | Reference | |||||||||
| TRP channel construct | Interactor construct | |||||||||
| TRP channel | Interactor | Method | Species | Region | Expression system | Species | Region | Expression system | ||
| TRPV1 |
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SNAPIN | Fusion protein-pull down assay | Rat | 1-414 | E. coli | Human | Full-length | In vitro translation | 15066994 |
| TRPV1 |
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Synaptotagmin 1 | Fusion protein-pull down assay | Rat | 1-414 | E. coli | Human | Full-length | In vitro translation | 15066994 |
| TRPV1 |
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Synaptotagmin 4 | Fusion protein-pull down assay | Rat | 1-414 | E. coli | Human | Full-length | In vitro translation | 15066994 |
(:
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
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top
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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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SNAPIN | Co-immunoprecipitation | HEK293 | Human | Full-length | Human | Full-length | 15066994 | |
| TRPV1 |
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Synaptotagmin 1 | Co-immunoprecipitation | HEK293 | Human | Full-length | Human | Full-length | 15066994 | |
| TRPV1 |
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Synaptotagmin 4 | Co-immunoprecipitation | HEK293 | Human | Full-length | Human | Full-length | 15066994 | |
| TRPV1 |
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Synaptotagmin 4 | Co-immunofluorescence staining | Rat dorsal root ganglion neuron | 15066994 | |||||
(:
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
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| Characterization
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| Binding region mapping | Stoichiometry | Affinity (Kd) | Reference | |||||||
| TRP channel | Interactor | |||||||||
| TRP channel | Interactor | Method | Species | Region | Species | Region | ||||
| TRPV1 |
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Synaptotagmin 4 | Fusion protein-pull down assay | Rat | 1-414 | Human | C2A domain | 15066994 | ||
(:
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)