J Biol Chem. 2012 May 11;287(20):16645-55. Epub 2012 Mar 27.

Calmodulin and S100A1 Protein Interact with N Terminus of TRPM3 Channel.External

Holakovska, B., Grycova, L., Jirku, M., Sulc, M., Bumba, L., Teisinger, J.,
["From the Institute of Physiology and."]
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: 22451665External
Arrow2 Screening Arrow2 Validation: In vitro validation Arrow2 Validation: In vivo validation Arrow2 Characterization Arrow2 Functional consequence Arrow2 top
Screening Toggle
  Experimental screening Non-experimental screening Reference
TRP channel construct Interactor source
TRP channel Interactor Method Species Region Species Organ/tissue Sample type
TRPM3 Link Calmodulin Inference Prediction 22451665
TRPM3 Link S100A10 Inference Prediction 22451665
(Link: click the arrow icon to show interactions only between the corresponding TRP channel and the interactor)
Arrow2 Screening Arrow2 Validation: In vitro validation Arrow2 Validation: In vivo validation Arrow2 Characterization Arrow2 Functional consequence Arrow2 top
Validation: In vitro validation Toggle
  Assay with recombinant proteins Reference
TRP channel construct Interactor construct
TRP channel Interactor Method Species Region Expression system Species Region Expression system
TRPM3 Link Calmodulin Fluorescence probe labeling Human 35-124 E. coli Not specified Not specified Not specified 22451665
TRPM3 Link Calmodulin Fluorescence probe labeling Human 291-382 E. coli Not specified Not specified Not specified 22451665
TRPM3 Link S100A10 Fluorescence probe labeling Human 35-124 E. coli Human Full-length E. coli 22451665
TRPM3 Link S100A10 Fluorescence probe labeling Human 291-382 E. coli Human Full-length E. coli 22451665
(Link: click the arrow icon to show interactions only between the corresponding TRP channel and the interactor)
Arrow2 Screening Arrow2 Validation: In vitro validation Arrow2 Validation: In vivo validation Arrow2 Characterization Arrow2 Functional consequence Arrow2 top
Characterization Toggle
  Binding region mapping Stoichiometry Affinity (Kd) Reference
TRP channel Interactor
TRP channel Interactor Method Species Region Species Region
TRPM3 Link Calmodulin Fluorescence probe labeling Human 291-382 Not specified Not determined 22451665
TRPM3 Link Calmodulin Fluorescence probe labeling Human 35-124 Not specified Not determined 22451665
TRPM3 Link S100A10 Fluorescence probe labeling Human 291-382 Human Not determined 22451665
TRPM3 Link S100A10 Fluorescence probe labeling Human 35-124 Human Not determined 22451665
(Link: click the arrow icon to show interactions only between the corresponding TRP channel and the interactor)
TRP / Interactor

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