J Proteome Res. 2008 Mar;7(3):979-89. doi: 10.1021/pr070496k. Epub 2008 Jan 19.

Analysis of TRPC3-interacting proteins by tandem mass spectrometry.External 2231691f894ba696de1310221b0a0dbbb31a7251e75115c265587c3d9d5f507c

Lockwich, T., Pant, J., Makusky, A., Jankowska-Stephens, E., Kowalak, J. A., Markey, S. P., Ambudkar, I. S.,
["Secretory Physiology Section, Molecular Physiology and Therapeutics Branch, NIDCR, National Institutes of Health, Bethesda, Maryland 20892, USA."]
Mammalian transient receptor potential canonical (TRPC) channels are a family of nonspecific cation channels that are activated in response to stimulation of phospholipase C (PLC)-dependent hydrolysis of the membrane lipid phosphatidylinositol 4,5-bisphosphate. Despite extensive studies, the mechanism(s) involved in regulation of mammalian TRPC channels remains unknown. Presence of various protein-interacting domains in TRPC channels have led to the suggestion that they associate with proteins that are involved in their function and regulation. This study was directed toward identifying the proteins associated with native TRPC3 using a shotgun proteomic approach. Anti-TRPC3 antibody was used to immunoprecipitate TRPC3 from solubilized rat brain crude membranes under conditions that allow retention of TRPC3 function. Proteins in the TRPC3 (using anti-TRPC3 antibody) and control (using rabbit IgG) immunoprecipitates were separated by SDS-PAGE, the gel was sectioned, and the resolved proteins were digested by trypsin in situ. After extraction of the peptides, the peptides were separated by HPLC and sequences derived by MS/MS. Analysis of the data revealed 64 specific TRPC3-associated proteins which can be grouped in terms of their cellular location and involvement in specific cellular function. Many of the proteins identified have been previously reported as TRPC3-regulatory proteins, such as IP3Rs and vesicle trafficking proteins. In addition, we report novel putative TRPC3-interacting proteins, including those involved in protein endocytosis and neuronal growth. To our knowledge, this is the first comprehensive proteomic analysis of a native TRPC channel. These data reveal potential TRPC3 regulatory proteins and provide novel insights of the mechanism(s) regulating TRPC3 channels as well as the possible cellular functions where the channel might be involved.
PMID: 18205297External 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
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 14-3-3 protein Ƣ/ơ Affinity purification-mass spectrometry Not used as a bait Rat Brain Crude membranes 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 14-3-3 protein ƥ Affinity purification-mass spectrometry Not used as a bait Rat Brain Crude membranes 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 14-3-3 protein Ƨ Affinity purification-mass spectrometry Not used as a bait Rat Brain Crude membranes 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 Aconitase Affinity purification-mass spectrometry Not used as a bait Rat Brain Crude membranes 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 ADT1 Affinity purification-mass spectrometry Not used as a bait Rat Brain Crude membranes 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 ALDOC Affinity purification-mass spectrometry Not used as a bait Rat Brain Crude membranes 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 Annexin 5 Affinity purification-mass spectrometry Not used as a bait Rat Brain Crude membranes 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 AP2A2 Affinity purification-mass spectrometry Not used as a bait Rat Brain Crude membranes 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 AP2B1 Affinity purification-mass spectrometry Not used as a bait Rat Brain Crude membranes 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 ATP synthase Ƣ Affinity purification-mass spectrometry Not used as a bait Rat Brain Crude membranes 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 Basp1 Affinity purification-mass spectrometry Not used as a bait Rat Brain Crude membranes 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 Caskin-1 Affinity purification-mass spectrometry Not used as a bait Rat Brain Crude membranes 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 Citrate synthase Affinity purification-mass spectrometry Not used as a bait Rat Brain Crude membranes 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 Clathrin-H1 Affinity purification-mass spectrometry Not used as a bait Rat Brain Crude membranes 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 Cytochrome c oxidase 2 Affinity purification-mass spectrometry Not used as a bait Rat Brain Crude membranes 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 Cytochrome c oxidase 5A Affinity purification-mass spectrometry Not used as a bait Rat Brain Crude membranes 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 DRP-2 Affinity purification-mass spectrometry Not used as a bait Rat Brain Crude membranes 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 Dynamin-1 heavy chain 1 Affinity purification-mass spectrometry Not used as a bait Rat Brain Crude membranes 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 Dynamin-1L Affinity purification-mass spectrometry Not used as a bait Rat Brain Crude membranes 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 Fibronectin Affinity purification-mass spectrometry Not used as a bait Rat Brain Crude membranes 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 Fumarase Affinity purification-mass spectrometry Not used as a bait Rat Brain Crude membranes 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 GLUD1 Affinity purification-mass spectrometry Not used as a bait Rat Brain Crude membranes 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 Gơq Affinity purification-mass spectrometry Not used as a bait Rat Brain Crude membranes 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 Gz-ơ Affinity purification-mass spectrometry Not used as a bait Rat Brain Crude membranes 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 Idh3B Affinity purification-mass spectrometry Not used as a bait Rat Brain Crude membranes 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 IP3R1 Affinity purification-mass spectrometry Not used as a bait Rat Brain Crude membranes 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 LETM1 Affinity purification-mass spectrometry Not used as a bait Rat Brain Crude membranes 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 MAP2 Affinity purification-mass spectrometry Not used as a bait Rat Brain Crude membranes 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 MDH1 Affinity purification-mass spectrometry Not used as a bait Rat Brain Crude membranes 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 MDH2 Affinity purification-mass spectrometry Not used as a bait Rat Brain Crude membranes 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 Myosin-10 Affinity purification-mass spectrometry Not used as a bait Rat Brain Crude membranes 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 NCAM1 Affinity purification-mass spectrometry Not used as a bait Rat Brain Crude membranes 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 Neurofascin Affinity purification-mass spectrometry Not used as a bait Rat Brain Crude membranes 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 Neuromodulin Affinity purification-mass spectrometry Not used as a bait Rat Brain Crude membranes 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 NKAơ1 Affinity purification-mass spectrometry Not used as a bait Rat Brain Crude membranes 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 NKAơ2 Affinity purification-mass spectrometry Not used as a bait Rat Brain Crude membranes 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 NKAơ3 Affinity purification-mass spectrometry Not used as a bait Rat Brain Crude membranes 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 ORP-150 Affinity purification-mass spectrometry Not used as a bait Rat Brain Crude membranes 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 p97/VCP Affinity purification-mass spectrometry Not used as a bait Rat Brain Crude membranes 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 PEBP1 Affinity purification-mass spectrometry Not used as a bait Rat Brain Crude membranes 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 PGK1 Affinity purification-mass spectrometry Not used as a bait Rat Brain Crude membranes 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 PMCA Affinity purification-mass spectrometry Not used as a bait Rat Brain Crude membranes 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 PMCA2 Affinity purification-mass spectrometry Not used as a bait Rat Brain Crude membranes 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 PMCA3 Affinity purification-mass spectrometry Not used as a bait Rat Brain Crude membranes 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 PMCA4 Affinity purification-mass spectrometry Not used as a bait Rat Brain Crude membranes 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 Pyruvate kinase M1/M2 Affinity purification-mass spectrometry Not used as a bait Rat Brain Crude membranes 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 RAP1A Affinity purification-mass spectrometry Not used as a bait Rat Brain Crude membranes 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 RAP1B Affinity purification-mass spectrometry Not used as a bait Rat Brain Crude membranes 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 SERCA2 Affinity purification-mass spectrometry Not used as a bait Rat Brain Crude membranes 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 Slc1a2 Affinity purification-mass spectrometry Not used as a bait Rat Brain Crude membranes 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 Slc1a3 Affinity purification-mass spectrometry Not used as a bait Rat Brain Crude membranes 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 SNAP25 Affinity purification-mass spectrometry Not used as a bait Rat Brain Crude membranes 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 SOD2 Affinity purification-mass spectrometry Not used as a bait Rat Brain Crude membranes 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 SV2A Affinity purification-mass spectrometry Not used as a bait Rat Brain Crude membranes 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 Synapsin1 Affinity purification-mass spectrometry Not used as a bait Rat Brain Crude membranes 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 Synaptotagmin 1 Affinity purification-mass spectrometry Not used as a bait Rat Brain Crude membranes 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 Syntaxin1a Affinity purification-mass spectrometry Not used as a bait Rat Brain Crude membranes 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 Tenascin-R Affinity purification-mass spectrometry Not used as a bait Rat Brain Crude membranes 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 UCH-L1 Affinity purification-mass spectrometry Not used as a bait Rat Brain Crude membranes 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 VAMP2 Affinity purification-mass spectrometry Not used as a bait Rat Brain Crude membranes 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 VAT2 Affinity purification-mass spectrometry Not used as a bait Rat Brain Crude membranes 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 VDAC1 Affinity purification-mass spectrometry Not used as a bait Rat Brain Crude membranes 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 VDAC3 Affinity purification-mass spectrometry Not used as a bait Rat Brain Crude membranes 18205297
(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
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 ADT1 Co-immunoprecipitation Rat brain lysates 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 Clathrin-H1 Co-immunoprecipitation Rat brain lysates 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 DRP-2 Co-immunoprecipitation Rat brain lysates 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 Dynamin-1 heavy chain 1 Co-immunoprecipitation Rat brain lysates 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 Gz-ơ Co-immunoprecipitation Rat brain lysates 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 IP3R1 Co-immunoprecipitation Rat brain lysates 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 Neuromodulin Co-immunoprecipitation Rat brain lysates 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 NKAơ1 Co-immunoprecipitation Rat brain lysates 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 PMCA Co-immunoprecipitation Rat brain lysates 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 SERCA2 Co-immunoprecipitation Rat brain lysates 18205297
TRPC3 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 Synaptotagmin 1 Co-immunoprecipitation Rat brain lysates 18205297
(Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4: click the arrow icon to show interactions only between the corresponding TRP channel and the interactor)
TRP / Interactor

To prevent spam comments, we use reCAPTCHA. Please type correct words into the following box.