J Biol Chem. 2012 May 11;287(20):16645-55. doi: 10.1074/jbc.M112.350686. Epub 2012 Mar 27.
Calmodulin and S100A1 protein interact with N terminus of TRPM3 channel.
Holakovska, B., Grycova, L., Jirku, M., Sulc, M., Bumba, L., Teisinger, J.,
["Institute of Physiology, Academy of Sciences of the Czech Republic, Videnska 1083, 142 20 Prague, Czech Republic."]
["Institute of Physiology, Academy of Sciences of the Czech Republic, Videnska 1083, 142 20 Prague, Czech Republic."]
Transient receptor potential melastatin 3 ion channel (TRPM3) belongs to the TRP family of cation-permeable ion channels involved in many important biological functions such as pain transduction, thermosensation, and mechanoregulation. The channel was reported to play an important role in Ca(2+) homeostasis, but its gating mechanisms, functions, and regulation are still under research. Utilizing biophysical and biochemical methods, we characterized two independent domains, Ala-35-Lys-124 and His-291-Gly-382, on the TRPM3 N terminus, responsible for interactions with the Ca(2+)-binding proteins calmodulin (CaM) and S100A1. We identified several positively charged residues within these domains as having a crucial impact on CaM/S100A1 binding. The data also suggest that the interaction is calcium-dependent. We also performed competition assays, which suggested that CaM and S100A1 are able to compete for the same binding sites within the TRPM3 N terminus. This is the first time that such an interaction has been shown for TRP family members.
PMID: 22451665
 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 | |||
| TRPM3 |
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Calmodulin | Inference | Prediction | 22451665 | |||||
| TRPM3 |
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S100A10 | Inference | Prediction | 22451665 | |||||
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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 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 | ||
| TRPM3 |
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Calmodulin | Fluorescence probe labeling | Human | 35-124 | E. coli | Not specified | Not specified | Not specified | 22451665 |
| TRPM3 |
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Calmodulin | Fluorescence probe labeling | Human | 291-382 | E. coli | Not specified | Not specified | Not specified | 22451665 |
| TRPM3 |
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S100A10 | Fluorescence probe labeling | Human | 35-124 | E. coli | Human | Full-length | E. coli | 22451665 |
| TRPM3 |
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S100A10 | Fluorescence probe labeling | Human | 291-382 | E. coli | Human | Full-length | E. coli | 22451665 |
(:
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
|
| Characterization
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|---|---|---|---|---|---|---|---|---|---|---|
| Binding region mapping | Stoichiometry | Affinity (Kd) | Reference | |||||||
| TRP channel | Interactor | |||||||||
| TRP channel | Interactor | Method | Species | Region | Species | Region | ||||
| TRPM3 |
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Calmodulin | Fluorescence probe labeling | Human | 291-382 | Not specified | Not determined | 22451665 | ||
| TRPM3 |
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Calmodulin | Fluorescence probe labeling | Human | 35-124 | Not specified | Not determined | 22451665 | ||
| TRPM3 |
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S100A10 | Fluorescence probe labeling | Human | 291-382 | Human | Not determined | 22451665 | ||
| TRPM3 |
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S100A10 | Fluorescence probe labeling | Human | 35-124 | Human | Not determined | 22451665 | ||
(:
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)