PMID- 28882890 OWN - NLM STAT- MEDLINE DCOM- 20171026 LR - 20210205 IS - 1083-351X (Electronic) IS - 0021-9258 (Print) IS - 0021-9258 (Linking) VI - 292 IP - 42 DP - 2017 Oct 20 TI - C-terminal phosphorylation of Na(V)1.5 impairs FGF13-dependent regulation of channel inactivation. PG - 17431-17448 LID - 10.1074/jbc.M117.787788 [doi] AB - Voltage-gated Na(+) (Na(V)) channels are key regulators of myocardial excitability, and Ca(2+)/calmodulin-dependent protein kinase II (CaMKII)-dependent alterations in Na(V)1.5 channel inactivation are emerging as a critical determinant of arrhythmias in heart failure. However, the global native phosphorylation pattern of Na(V)1.5 subunits associated with these arrhythmogenic disorders and the associated channel regulatory defects remain unknown. Here, we undertook phosphoproteomic analyses to identify and quantify in situ the phosphorylation sites in the Na(V)1.5 proteins purified from adult WT and failing CaMKIIdelta(c)-overexpressing (CaMKIIdelta(c)-Tg) mouse ventricles. Of 19 native Na(V)1.5 phosphorylation sites identified, two C-terminal phosphoserines at positions 1938 and 1989 showed increased phosphorylation in the CaMKIIdelta(c)-Tg compared with the WT ventricles. We then tested the hypothesis that phosphorylation at these two sites impairs fibroblast growth factor 13 (FGF13)-dependent regulation of Na(V)1.5 channel inactivation. Whole-cell voltage-clamp analyses in HEK293 cells demonstrated that FGF13 increases Na(V)1.5 channel availability and decreases late Na(+) current, two effects that were abrogated with Na(V)1.5 mutants mimicking phosphorylation at both sites. Additional co-immunoprecipitation experiments revealed that FGF13 potentiates the binding of calmodulin to Na(V)1.5 and that phosphomimetic mutations at both sites decrease the interaction of FGF13 and, consequently, of calmodulin with Na(V)1.5. Together, we have identified two novel native phosphorylation sites in the C terminus of Na(V)1.5 that impair FGF13-dependent regulation of channel inactivation and may contribute to CaMKIIdelta(c)-dependent arrhythmogenic disorders in failing hearts. CI - (c) 2017 by The American Society for Biochemistry and Molecular Biology, Inc. FAU - Burel, Sophie AU - Burel S AD - From the l'Institut du Thorax, INSERM, CNRS, UNIV Nantes, Nantes 44007, France. FAU - Coyan, Fabien C AU - Coyan FC AD - From the l'Institut du Thorax, INSERM, CNRS, UNIV Nantes, Nantes 44007, France. FAU - Lorenzini, Maxime AU - Lorenzini M AD - From the l'Institut du Thorax, INSERM, CNRS, UNIV Nantes, Nantes 44007, France. FAU - Meyer, Matthew R AU - Meyer MR AD - the Departments of Medicine. FAU - Lichti, Cheryl F AU - Lichti CF AD - the Department of Pharmacology and Toxicology, University of Texas Medical Branch, Galveston, Texas 77555. FAU - Brown, Joan H AU - Brown JH AD - the Department of Pharmacology, University of California at San Diego, La Jolla, California 92093-0636, and. FAU - Loussouarn, Gildas AU - Loussouarn G AD - From the l'Institut du Thorax, INSERM, CNRS, UNIV Nantes, Nantes 44007, France. FAU - Charpentier, Flavien AU - Charpentier F AD - From the l'Institut du Thorax, INSERM, CNRS, UNIV Nantes, Nantes 44007, France. FAU - Nerbonne, Jeanne M AU - Nerbonne JM AD - Developmental Biology. AD - Internal Medicine, and. FAU - Townsend, R Reid AU - Townsend RR AD - Internal Medicine, and. AD - Cell Biology and Physiology, Washington University Medical School, St. Louis, Missouri 63110. FAU - Maier, Lars S AU - Maier LS AD - the Department of Internal Medicine II, University Heart Center, University Hospital Regensburg, D-93042 Regensburg, Germany. FAU - Marionneau, Celine AU - Marionneau C AD - From the l'Institut du Thorax, INSERM, CNRS, UNIV Nantes, Nantes 44007, France, celine.marionneau@univ-nantes.fr. LA - eng GR - P01 HL080101/HL/NHLBI NIH HHS/United States GR - P41 GM103422/GM/NIGMS NIH HHS/United States GR - UL1 TR000448/TR/NCATS NIH HHS/United States GR - UL1 TR002345/TR/NCATS NIH HHS/United States GR - R01 HL034161/HL/NHLBI NIH HHS/United States PT - Journal Article DEP - 20170907 PL - United States TA - J Biol Chem JT - The Journal of biological chemistry JID - 2985121R RN - 0 (NAV1.5 Voltage-Gated Sodium Channel) RN - 0 (SCN5A protein, human) RN - 0 (Scn5a protein, mouse) RN - 330E71H5Q4 (fibroblast growth factor 13) RN - 62031-54-3 (Fibroblast Growth Factors) RN - EC 2.7.11.17 (Calcium-Calmodulin-Dependent Protein Kinase Type 2) SB - IM MH - Amino Acid Substitution MH - Animals MH - Calcium-Calmodulin-Dependent Protein Kinase Type 2/genetics/metabolism MH - Fibroblast Growth Factors/genetics/*metabolism MH - HEK293 Cells MH - Heart Failure/genetics/*metabolism MH - Humans MH - *Ion Channel Gating MH - Mice MH - Mice, Transgenic MH - Mutation, Missense MH - NAV1.5 Voltage-Gated Sodium Channel/genetics/*metabolism MH - Phosphorylation PMC - PMC5655519 OTO - NOTNLM OT - Ca2+/calmodulin-dependent protein kinase II (CaMKII) OT - FGF13 OT - Nav1.5 OT - calmodulin (CaM) OT - channel inactivation OT - heart OT - phosphoproteomics OT - phosphorylation OT - sodium channel COIS- The authors declare that they have no conflicts of interest with the contents of this article EDAT- 2017/09/09 06:00 MHDA- 2017/10/27 06:00 PMCR- 2018/10/20 CRDT- 2017/09/09 06:00 PHST- 2017/03/22 00:00 [received] PHST- 2017/08/23 00:00 [revised] PHST- 2017/09/09 06:00 [pubmed] PHST- 2017/10/27 06:00 [medline] PHST- 2017/09/09 06:00 [entrez] PHST- 2018/10/20 00:00 [pmc-release] AID - S0021-9258(20)33900-4 [pii] AID - M117.787788 [pii] AID - 10.1074/jbc.M117.787788 [doi] PST - ppublish SO - J Biol Chem. 2017 Oct 20;292(42):17431-17448. doi: 10.1074/jbc.M117.787788. Epub 2017 Sep 7.