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.External 2231691f894ba696de1310221b0a0dbbb31a7251e75115c265587c3d9d5f507c

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."]
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: 17074976External 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 Catenin-ơ2 Yeast two-hybrid Rat 1-432 Human Fetal brain cDNA library 17074976
TRPV1 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 Catenin-Ƥ2 Yeast two-hybrid Rat 1-432 Human Fetal brain cDNA library 17074976
TRPV1 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 CDK12 Yeast two-hybrid Rat 1-432 Human Fetal brain cDNA library 17074976
TRPV1 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 CHERP Yeast two-hybrid Rat 1-432 Human Fetal brain cDNA library 17074976
TRPV1 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 Contactin-1 Yeast two-hybrid Rat 1-432 Human Fetal brain cDNA library 17074976
TRPV1 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 DAP-150 Yeast two-hybrid Rat 1-432 Human Fetal brain cDNA library 17074976
TRPV1 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 DAZAP2 Yeast two-hybrid Rat 1-432 Human Fetal brain cDNA library 17074976
TRPV1 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 GCN5 Yeast two-hybrid Rat 1-432 Human Fetal brain cDNA library 17074976
TRPV1 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 HAX-1 Yeast two-hybrid Rat 1-432 Human Fetal brain cDNA library 17074976
TRPV1 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 ITM2C Yeast two-hybrid Rat 1-432 Human Fetal brain cDNA library 17074976
TRPV1 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 JMJD2C Yeast two-hybrid Rat 1-432 Human Fetal brain cDNA library 17074976
TRPV1 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 KIF2A Yeast two-hybrid Rat 1-432 Human Fetal brain cDNA library 17074976
TRPV1 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 KIF3B Yeast two-hybrid Rat 1-432 Human Fetal brain cDNA library 17074976
TRPV1 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 KPNƢ1 Yeast two-hybrid Rat 1-432 Human Fetal brain cDNA library 17074976
TRPV1 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 Neuronatin Yeast two-hybrid Rat 1-432 Human Fetal brain cDNA library 17074976
TRPV1 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 NKX2.2 Yeast two-hybrid Rat 1-432 Human Fetal brain cDNA library 17074976
TRPV1 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 NOMO1 Yeast two-hybrid Rat 1-432 Human Fetal brain cDNA library 17074976
TRPV1 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 PEX10 Yeast two-hybrid Rat 1-432 Human Fetal brain cDNA library 17074976
TRPV1 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 PI3K-p85Ƣ Yeast two-hybrid Rat 1-432 Human Fetal brain cDNA library 17074976
TRPV1 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 PI3K-p85ơ Inference Prediction 17074976
TRPV1 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 PLAGL1 Yeast two-hybrid Rat 1-432 Human Fetal brain cDNA library 17074976
TRPV1 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 POLR2G Yeast two-hybrid Rat 1-432 Human Fetal brain cDNA library 17074976
TRPV1 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 POLR3D Yeast two-hybrid Rat 1-432 Human Fetal brain cDNA library 17074976
TRPV1 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 RAB26 Yeast two-hybrid Rat 1-432 Human Fetal brain cDNA library 17074976
TRPV1 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 RANBP9 Yeast two-hybrid Rat 1-432 Human Fetal brain cDNA library 17074976
TRPV1 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 RPL12 Yeast two-hybrid Rat 1-432 Human Fetal brain cDNA library 17074976
TRPV1 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 SGT Yeast two-hybrid Rat 1-432 Human Fetal brain cDNA library 17074976
TRPV1 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 SMA3 Yeast two-hybrid Rat 1-432 Human Fetal brain cDNA library 17074976
TRPV1 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 SNAPIN Yeast two-hybrid Rat 1-432 Human Fetal brain cDNA library 17074976
TRPV1 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 TGS1 Yeast two-hybrid Rat 1-432 Human Fetal brain cDNA library 17074976
TRPV1 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 Thrombospondin-3 Yeast two-hybrid Rat 1-432 Human Fetal brain cDNA library 17074976
TRPV1 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 TMEM115 Yeast two-hybrid Rat 1-432 Human Fetal brain cDNA library 17074976
TRPV1 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 UBP5 Yeast two-hybrid Rat 1-432 Human Fetal brain cDNA library 17074976
TRPV1 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 ZBTB17 Yeast two-hybrid Rat 1-432 Human Fetal brain cDNA library 17074976
(Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4: click the arrow icon to show interactions only between the corresponding TRP channel and the interactor)
Validation: In vitro validation Toggle 893349bafcc528f8346c51dc3420151d67b0126b2c122dd1017121c03fa0f69b
  Assay with recombinant proteins Reference
TRP channel construct Interactor construct
TRP channel Interactor Method Species Region Expression system Species Region Expression system
TRPV1 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 PI3K-p85ơ Fusion protein-pull down assay Rat Full-length HEK293 lysates Not specified Full-length E. coli 17074976
(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 PI3K-p85Ƣ Co-immunoprecipitation HEK293 Rat Full-length Not used 17074976
TRPV1 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 PI3K-p85Ƣ Co-immunoprecipitation Mouse dorsal root ganglion cell 17074976
(Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4: click the arrow icon to show interactions only between the corresponding TRP channel and the interactor)
Characterization Toggle 893349bafcc528f8346c51dc3420151d67b0126b2c122dd1017121c03fa0f69b
  Binding region mapping Stoichiometry Affinity (Kd) Reference
TRP channel Interactor
TRP channel Interactor Method Species Region Species Region
TRPV1 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 PI3K-p85ơ Fusion protein-pull down assay Rat N-terminus Not specified SH2 domain 17074976
(Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4: click the arrow icon to show interactions only between the corresponding TRP channel and the interactor)
TRP / Interactor

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