J Physiol. 2006 Apr 1;572(Pt 1):165-72. Epub 2006 Feb 9.
Calcium-sensing mechanism in TRPC5 channels contributing to retardation of neurite outgrowth.
Hui, H., McHugh, D., Hannan, M., Zeng, F., Xu, S. Z., Khan, S. U., Levenson, R., Beech, D. J., Weiss, J. L.,
["Institute of Membrane and Systems Biology, Garstang Building, University of Leeds, Leeds LS2 9JT, UK."]
["Institute of Membrane and Systems Biology, Garstang Building, University of Leeds, Leeds LS2 9JT, UK."]
The calcium- and sodium-permeable transient receptor potential channel TRPC5 has an inhibitory role in neuronal outgrowth but the mechanisms governing its activity are poorly understood. Here we propose a mechanism involving the neuronal calcium sensor-1 (NCS-1) protein. Inhibitory mutants of TRPC5 and NCS-1 enhance neurite outgrowth similarly. Mutant NCS-1 does not inhibit surface-expression of TRPC5 but generally suppresses channel activity, irrespective of whether it is evoked by carbachol, store depletion, lanthanides or elevated intracellular calcium. NCS-1 and TRPC5 are in the same protein complex in rat brain and NCS-1 directly binds to the TRPC5 C-terminus. The data suggest protein-protein interaction between NCS-1 and TRPC5, and involvement of this protein complex in retardation of neurite outgrowth.
PMID: 16469785
 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 | |||
| TRPC5 |
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NCS-1 | Inference | Prediction | 16469785 | |||||
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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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| 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 | ||
| TRPC5 |
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NCS-1 | Yeast two-hybrid | Human | 619-973 | Yeast | Human | Full-length | Yeast | 16469785 |
| TRPC5 |
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NCS-1 | Fusion protein-pull down assay | Human | 619-973 | E. coli | Human | Full-length | E. coli | 16469785 |
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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 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 | ||||
| TRPC5 |
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NCS-1 | Co-immunoprecipitation | HEK293 | Human | Full-length | Human | Full-length | 16469785 | |
| TRPC5 |
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NCS-1 | Co-immunoprecipitation | Rat brain membrane lysates | 16469785 | |||||
(:
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)
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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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| Characterization
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|---|---|---|---|---|---|---|---|---|---|---|
| Binding region mapping | Stoichiometry | Affinity (Kd) | Reference | |||||||
| TRP channel | Interactor | |||||||||
| TRP channel | Interactor | Method | Species | Region | Species | Region | ||||
| TRPC5 |
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NCS-1 | Yeast two-hybrid | Human | 619-803 | Human | Not determined | 16469785 | ||
| TRPC5 |
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NCS-1 | Yeast two-hybrid | Human | 803-973 | Human | Not determined | 16469785 | ||
| TRPC5 |
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NCS-1 | Fusion protein-pull down assay | Human | 619-803 | Human | Not determined | 16469785 | ||
| TRPC5 |
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NCS-1 | Fusion protein-pull down assay | Human | 803-973 | Human | Not determined | 16469785 | ||
(:
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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| Functional consequence
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| TRP channel | Interactor | Method | Post-translational modification | Subcellular trafficking | Activity | Reference | ||||||
| TRPC5 |
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NCS-1 | Calcium measurement | Activation | 16469785 | |||||||
| TRPC5 |
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NCS-1 | Patch clamp | Activation | 16469785 | |||||||
(:
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)