J Biol Chem. 2000 Dec 1;275(48):37559-64.
Association of mammalian trp4 and phospholipase C isozymes with a PDZ domain-containing protein, NHERF.
Tang, Y., Tang, J., Chen, Z., Trost, C., Flockerzi, V., Li, M., Ramesh, V., Zhu, M. X.,
["Neurobiotechnology Center and Department of Neuroscience, Ohio State University, Columbus, Ohio 43210, USA."]
["Neurobiotechnology Center and Department of Neuroscience, Ohio State University, Columbus, Ohio 43210, USA."]
Mammalian homologues of Drosophila Trp have been implicated to form channels that are activated following the depletion of Ca(2+) from internal stores. Recent studies indicate that actin redistribution is required for the activation of these channels. Here we show that murine Trp4 and Trp5, as well as phospholipase C beta1 and beta2 interact with the first PDZ domain of NHERF, regulatory factor of the Na(+)/H(+) exchanger. We demonstrated the association of Trp4 and phospholipase C-beta1 with NHERF in vivo in an HEK293 cell line expressing Trp4 and in adult mouse brain by immuno-coprecipitation. NHERF is a two PDZ domain-containing protein that associates with the actin cytoskeleton via interactions with members of ezrin/radixin/moesin family. Thus, store-operated channels involving Trp4 and Trp5 can form signaling complexes with phospholipase C isozymes via interactions with NHERF and thereby linking the lipase and the channels to the actin cytoskeleton. The interaction with the PDZ protein may constitute an important mechanism for distribution and regulation of store-operated channels.
PMID: 10980202

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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 | |||
TRPC4 |
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NHERF1 | Inference | Prediction | 10980202 | |||||
TRPC4 |
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PLCƢ1 | Inference | Prediction | 10980202 | |||||
TRPC5 |
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NHERF1 | Inference | Prediction | 10980202 |
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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 | ||
TRPC4 |
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NHERF1 | Fusion protein-pull down assay | Mouse | Full-length | In vitro translation | Mouse | 1-107 | E. coli | 10980202 |
TRPC5 |
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NHERF1 | Fusion protein-pull down assay | Mouse | Full-length | In vitro translation | Mouse | 1-107 | E. coli | 10980202 |
(
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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 | ||||
TRPC4 |
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NHERF1 | Co-immunoprecipitation | HEK293 | Mouse | Full-length | Not used | 10980202 | ||
TRPC4 |
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NHERF1 | Co-immunoprecipitation | Mouse brain tissue | 10980202 | |||||
TRPC4 |
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PLCƢ1 | Co-immunoprecipitation | HEK293 | Mouse | Full-length | Not used | 10980202 | ||
TRPC4 |
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PLCƢ1 | Co-immunoprecipitation | Mouse brain tissue | 10980202 |
(
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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 | ||||
TRPC4 |
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NHERF1 | Fusion protein-pull down assay | Mouse | 688-974 | Mouse | 1-107 | 10980202 | ||
TRPC5 |
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NHERF1 | Fusion protein-pull down assay | Mouse | 648-975 | Mouse | 1-107 | 10980202 |
(
:
click the arrow icon to show interactions only between the corresponding TRP channel and the interactor)
