title: Functional characterisation of a TRPM2 orthologue from the sea anemone Nematostella vectensis in human cells.
abstract: The human non-selective cation channel TRPM2 represents a mediator of apoptosis triggered by oxidative stress. The principal agonist ADP-ribose binds to the cytosolic domain of TRPM2, which is homologous to the human ADP-ribose pyrophosphatase NUDT9. To further elucidate the structure-function relationship of this channel, we characterised a TRPM2 orthologue from the cnidarian Nematostella vectensis, after its expression in a human cell line. This far distant relative shows only 31% total sequence similarity to hTRPM2, while its C-terminal domain has a greater resemblance to the NUDT9 enzyme. Current through nvTRPM2 was induced by ADPR, with a more pronounced sensitivity and faster kinetics than in hTRPM2. In contrast to hTRPM2, there was no response to H2O2 and hardly any modulatory effect by intracellular Ca(2+). The deletion of a stretch of 15 residues from the NUDT9 domain of nvTRPM2, which is absent in hTRPM2, did not change the response to ADPR but enabled activation of the channel by H2O2 and increased the effects of intracellular Ca(2+). These findings shed new light on the evolution of TRPM2 and establish nvTRPM2 as a promising tool to decipher its complex gating mechanisms.
authors.affiliation: Institute of Physiology, Medical Faculty, RWTH Aachen, D52057 Aachen, Germany.
authors.forename: Frank J P
authors.initials: FJ
authors.lastname: Kühn
authors.affiliation: Institute of Physiology, Medical Faculty, RWTH Aachen, D52057 Aachen, Germany.
authors.forename: Cornelia
authors.initials: C
authors.lastname: Kühn
authors.affiliation: Institute of Physiology, Medical Faculty, RWTH Aachen, D52057 Aachen, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Lückhoff
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002460:Cell Line; D006801:Humans; D018384:Oxidative Stress; D018408:Patch-Clamp Techniques; D011755:Pyrophosphatases / Q000235:genetics; D012615:Sea Anemones / Q000235:genetics*; D050053:TRPM Cation Channels / Q000235:genetics* / Q000378:metabolism
chemical_list: D050053:TRPM Cation Channels; C121934:TRPM2 protein, human; C429463:NUDT9; D011755:Pyrophosphatases; D002118:Calcium