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Updating In Vivo and In Vitro Phosphorylation and Methylation Sites of Voltage‐Gated Kv7.2 Potassium Channels
Author(s) -
Erdem Fatma Asli,
Salzer Isabella,
Heo Seok,
Chen WeiQiang,
Jung Gangsoo,
Lubec Gert,
Boehm Stefan,
Yang JaeWon
Publication year - 2017
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.201700015
Subject(s) - in vivo , in vitro , methylation , potassium channel , phosphorylation , microbiology and biotechnology , chemistry , potassium , voltage gated potassium channel , voltage gated ion channel , biology , computational biology , biochemistry , biophysics , genetics , ion channel , dna , receptor , organic chemistry
Voltage‐gated Kv7.2 potassium channels regulate neuronal excitability. The gating of these channels is tightly controlled by various mediators and neurotransmitters acting via G protein‐coupled receptors; the underlying signaling cascades involve phosphatidylinositol‐4,5‐bisphosphate (PIP 2 ), Ca 2+ /calmodulin, and phosphorylation. Recent studies found that the PIP 2 sensitivity of Kv7.2 channels is affected by two posttranslational modifications, phosphorylation and methylation, harboured within putative PIP 2 ‐binding domains. In this study, we updated phosphorylation and methylation sites in Kv7.2 either heterologously expressed in mammalian cells or as GST‐fusion proteins exposed to recombinant protein kinases by using LC–MS/MS. In vitro kinase assays revealed that CDK5, protein kinase C (PKC) alpha, PKA, p38 MAPK, CamKIIα, and GSK3β could mediate phosphorylation. Taken together, we provided a comprehensive map of phosphorylation and methylation in Kv7.2 within protein–protein and protein–lipid interaction domains. This may help to interpret the functional roles of individual PTM sites in Kv7.2 channels. All MS data are available via ProteomeXchange with the identifier PXD005567.