J Gen Physiol. 2006 Nov;128(5):509-22.
Phosphoinositide 3-kinase binds to TRPV1 and mediates NGF-stimulated TRPV1 trafficking to the plasma membrane.
Stein, A. T., Ufret-Vincenty, C. A., Hua, L., Santana, L. F., Gordon, S. E.,
["Graduate Program in Neurobiology and Behavior, University of Washington, Seattle, WA 98195, USA."]
["Graduate Program in Neurobiology and Behavior, University of Washington, Seattle, WA 98195, USA."]
Sensitization of the pain-transducing ion channel TRPV1 underlies thermal hyperalgesia by proalgesic agents such as nerve growth factor (NGF). The currently accepted model is that the NGF-mediated increase in TRPV1 function during hyperalgesia utilizes activation of phospholipase C (PLC) to cleave PIP2, proposed to tonically inhibit TRPV1. In this study, we tested the PLC model and found two lines of evidence that directly challenge its validity: (1) polylysine, a cationic phosphoinositide sequestering agent, inhibited TRPV1 instead of potentiating it, and (2) direct application of PIP2 to inside-out excised patches dramatically potentiated TRPV1. Furthermore, we show four types of experiments indicating that PI3K is physically and functionally coupled to TRPV1: (1) the p85beta subunit of PI3K interacted with the N-terminal region of TRPV1 in yeast 2-hybrid experiments, (2) PI3K-p85beta coimmunoprecipitated with TRPV1 from both HEK293 cells and dorsal root ganglia (DRG) neurons, (3) TRPV1 interacted with recombinant PI3K-p85 in vitro, and (4) wortmannin, a specific inhibitor of PI3K, completely abolished NGF-mediated sensitization in acutely dissociated DRG neurons. Finally, simultaneous electrophysiological and total internal reflection fluorescence (TIRF) microscopy recordings demonstrate that NGF increased the number of channels in the plasma membrane. We propose a new model for NGF-mediated hyperalgesia in which physical coupling of TRPV1 and PI3K in a signal transduction complex facilitates trafficking of TRPV1 to the plasma membrane.
PMID: 17074976

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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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Catenin-ơ2 | Yeast two-hybrid | Rat | 1-432 | Human | Fetal brain | cDNA library | 17074976 | |
TRPV1 |
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Catenin-Ƥ2 | Yeast two-hybrid | Rat | 1-432 | Human | Fetal brain | cDNA library | 17074976 | |
TRPV1 |
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CDK12 | Yeast two-hybrid | Rat | 1-432 | Human | Fetal brain | cDNA library | 17074976 | |
TRPV1 |
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CHERP | Yeast two-hybrid | Rat | 1-432 | Human | Fetal brain | cDNA library | 17074976 | |
TRPV1 |
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Contactin-1 | Yeast two-hybrid | Rat | 1-432 | Human | Fetal brain | cDNA library | 17074976 | |
TRPV1 |
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DAP-150 | Yeast two-hybrid | Rat | 1-432 | Human | Fetal brain | cDNA library | 17074976 | |
TRPV1 |
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DAZAP2 | Yeast two-hybrid | Rat | 1-432 | Human | Fetal brain | cDNA library | 17074976 | |
TRPV1 |
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GCN5 | Yeast two-hybrid | Rat | 1-432 | Human | Fetal brain | cDNA library | 17074976 | |
TRPV1 |
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HAX-1 | Yeast two-hybrid | Rat | 1-432 | Human | Fetal brain | cDNA library | 17074976 | |
TRPV1 |
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ITM2C | Yeast two-hybrid | Rat | 1-432 | Human | Fetal brain | cDNA library | 17074976 | |
TRPV1 |
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JMJD2C | Yeast two-hybrid | Rat | 1-432 | Human | Fetal brain | cDNA library | 17074976 | |
TRPV1 |
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KIF2A | Yeast two-hybrid | Rat | 1-432 | Human | Fetal brain | cDNA library | 17074976 | |
TRPV1 |
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KIF3B | Yeast two-hybrid | Rat | 1-432 | Human | Fetal brain | cDNA library | 17074976 | |
TRPV1 |
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KPNƢ1 | Yeast two-hybrid | Rat | 1-432 | Human | Fetal brain | cDNA library | 17074976 | |
TRPV1 |
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Neuronatin | Yeast two-hybrid | Rat | 1-432 | Human | Fetal brain | cDNA library | 17074976 | |
TRPV1 |
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NKX2.2 | Yeast two-hybrid | Rat | 1-432 | Human | Fetal brain | cDNA library | 17074976 | |
TRPV1 |
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NOMO1 | Yeast two-hybrid | Rat | 1-432 | Human | Fetal brain | cDNA library | 17074976 | |
TRPV1 |
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PEX10 | Yeast two-hybrid | Rat | 1-432 | Human | Fetal brain | cDNA library | 17074976 | |
TRPV1 |
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PI3K-p85Ƣ | Yeast two-hybrid | Rat | 1-432 | Human | Fetal brain | cDNA library | 17074976 | |
TRPV1 |
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PI3K-p85ơ | Inference | Prediction | 17074976 | |||||
TRPV1 |
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PLAGL1 | Yeast two-hybrid | Rat | 1-432 | Human | Fetal brain | cDNA library | 17074976 | |
TRPV1 |
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POLR2G | Yeast two-hybrid | Rat | 1-432 | Human | Fetal brain | cDNA library | 17074976 | |
TRPV1 |
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POLR3D | Yeast two-hybrid | Rat | 1-432 | Human | Fetal brain | cDNA library | 17074976 | |
TRPV1 |
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RAB26 | Yeast two-hybrid | Rat | 1-432 | Human | Fetal brain | cDNA library | 17074976 | |
TRPV1 |
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RANBP9 | Yeast two-hybrid | Rat | 1-432 | Human | Fetal brain | cDNA library | 17074976 | |
TRPV1 |
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RPL12 | Yeast two-hybrid | Rat | 1-432 | Human | Fetal brain | cDNA library | 17074976 | |
TRPV1 |
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SGT | Yeast two-hybrid | Rat | 1-432 | Human | Fetal brain | cDNA library | 17074976 | |
TRPV1 |
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SMA3 | Yeast two-hybrid | Rat | 1-432 | Human | Fetal brain | cDNA library | 17074976 | |
TRPV1 |
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SNAPIN | Yeast two-hybrid | Rat | 1-432 | Human | Fetal brain | cDNA library | 17074976 | |
TRPV1 |
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TGS1 | Yeast two-hybrid | Rat | 1-432 | Human | Fetal brain | cDNA library | 17074976 | |
TRPV1 |
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Thrombospondin-3 | Yeast two-hybrid | Rat | 1-432 | Human | Fetal brain | cDNA library | 17074976 | |
TRPV1 |
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TMEM115 | Yeast two-hybrid | Rat | 1-432 | Human | Fetal brain | cDNA library | 17074976 | |
TRPV1 |
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UBP5 | Yeast two-hybrid | Rat | 1-432 | Human | Fetal brain | cDNA library | 17074976 | |
TRPV1 |
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ZBTB17 | Yeast two-hybrid | Rat | 1-432 | Human | Fetal brain | cDNA library | 17074976 |
(
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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 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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PI3K-p85ơ | Fusion protein-pull down assay | Rat | Full-length | HEK293 lysates | Not specified | Full-length | E. coli | 17074976 |
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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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PI3K-p85Ƣ | Co-immunoprecipitation | HEK293 | Rat | Full-length | Not used | 17074976 | ||
TRPV1 |
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PI3K-p85Ƣ | Co-immunoprecipitation | Mouse dorsal root ganglion cell | 17074976 |
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click the arrow icon to show interactions only between the corresponding TRP channel and the interactor)

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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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PI3K-p85ơ | Fusion protein-pull down assay | Rat | N-terminus | Not specified | SH2 domain | 17074976 |
(
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click the arrow icon to show interactions only between the corresponding TRP channel and the interactor)
