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• W2056846690 endingPage "e97854" @default.
• W2056846690 startingPage "e97854" @default.
• W2056846690 abstract "The cytoplasmic loop between the second and third transmembrane segments is pivotal in the regulation of TRESK (TWIK-related spinal cord K+ channel, K2P18.1, KCNK18). Calcineurin binds to this region and activates the channel by dephosphorylation in response to the calcium signal. Phosphorylation-dependent anchorage of 14-3-3 adaptor protein also modulates TRESK at this location. In the present study, we identified molecular interacting partners of the intracellular loop. By an affinity chromatography approach using the cytoplasmic loop as bait, we have verified the specific association of calcineurin and 14-3-3 to the channel. In addition to these known interacting proteins, we observed substantial binding of tubulin to the intracellular loop. Successive truncation of the polypeptide and pull-down experiments from mouse brain cytosol narrowed down the region sufficient for the binding of tubulin to a 16 amino acid sequence: LVLGRLSYSIISNLDE. The first six residues of this sequence are similar to the previously reported tubulin-binding region of P2X2 purinergic receptor. The tubulin-binding site of TRESK is located close to the protein kinase A (PKA)-dependent 14-3-3-docking motif of the channel. We provide experimental evidence suggesting that 14-3-3 competes with tubulin for the binding to the cytoplasmic loop of TRESK. It is intriguing that the 16 amino acid tubulin-binding sequence includes the serines, which were previously shown to be phosphorylated by microtubule-affinity regulating kinases (MARK kinases) and contribute to channel inhibition. Although tubulin binds to TRESK in vitro, it remains to be established whether the two proteins also interact in the living cell." @default.
• W2056846690 created "2016-06-24" @default.
• W2056846690 creator A5025162460 @default.
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• W2056846690 creator A5059931044 @default.
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• W2056846690 date "2014-05-15" @default.
• W2056846690 modified "2023-09-30" @default.
• W2056846690 title "Tubulin Binds to the Cytoplasmic Loop of TRESK Background K+ Channel In Vitro" @default.
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• W2056846690 doi "https://doi.org/10.1371/journal.pone.0097854" @default.
• W2056846690 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/4022642" @default.
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• W2056846690 hasPublicationYear "2014" @default.
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