JAK2 regulates Nav1.6 channel function via FGF14
Action Potentials
/ drug effects
Amino Acid Sequence
Animals
Fibroblast Growth Factors
/ chemistry
HEK293 Cells
High-Throughput Screening Assays
Hippocampus
/ cytology
Humans
Janus Kinase 2
/ metabolism
Mice, Inbred C57BL
NAV1.6 Voltage-Gated Sodium Channel
/ metabolism
Phosphorylation
/ drug effects
Phosphotyrosine
/ metabolism
Protein Binding
/ drug effects
Protein Kinase Inhibitors
/ analysis
Protein Multimerization
/ drug effects
Pyramidal Cells
/ drug effects
Reproducibility of Results
src-Family Kinases
/ metabolism
FGF14
JAK2
Signaling
Sodium channels
Tyrosine kinases
Journal
Biochimica et biophysica acta. Molecular cell research
ISSN: 1879-2596
Titre abrégé: Biochim Biophys Acta Mol Cell Res
Pays: Netherlands
ID NLM: 101731731
Informations de publication
Date de publication:
10 2020
10 2020
Historique:
received:
10
03
2020
revised:
18
06
2020
accepted:
20
06
2020
pubmed:
1
7
2020
medline:
15
12
2020
entrez:
30
6
2020
Statut:
ppublish
Résumé
Protein interactions between voltage-gated sodium (Nav) channels and accessory proteins play an essential role in neuronal firing and plasticity. However, a surprisingly limited number of kinases have been identified as regulators of these molecular complexes. We hypothesized that numerous as-of-yet unidentified kinases indirectly regulate the Nav channel via modulation of the intracellular fibroblast growth factor 14 (FGF14), an accessory protein with numerous unexplored phosphomotifs and required for channel function in neurons. Here we present results from an in-cell high-throughput screening (HTS) against the FGF14: Nav1.6 complex using >3000 diverse compounds targeting an extensive range of signaling pathways. Regulation by top kinase targets was then explored using in vitro phosphorylation, biophysics, mass-spectrometry and patch-clamp electrophysiology. Compounds targeting Janus kinase 2 (JAK2) were over-represented among HTS hits. Phosphomotif scans supported by mass spectrometry revealed FGF14 These studies point toward a novel mechanism by which levels of JAK2 in neurons could directly influence firing and plasticity by controlling the FGF14 dimerization equilibrium, and thereby the availability of monomeric species for interaction with Nav1.6.
Sections du résumé
BACKGROUND
Protein interactions between voltage-gated sodium (Nav) channels and accessory proteins play an essential role in neuronal firing and plasticity. However, a surprisingly limited number of kinases have been identified as regulators of these molecular complexes. We hypothesized that numerous as-of-yet unidentified kinases indirectly regulate the Nav channel via modulation of the intracellular fibroblast growth factor 14 (FGF14), an accessory protein with numerous unexplored phosphomotifs and required for channel function in neurons.
METHODS
Here we present results from an in-cell high-throughput screening (HTS) against the FGF14: Nav1.6 complex using >3000 diverse compounds targeting an extensive range of signaling pathways. Regulation by top kinase targets was then explored using in vitro phosphorylation, biophysics, mass-spectrometry and patch-clamp electrophysiology.
RESULTS
Compounds targeting Janus kinase 2 (JAK2) were over-represented among HTS hits. Phosphomotif scans supported by mass spectrometry revealed FGF14
CONCLUSIONS
These studies point toward a novel mechanism by which levels of JAK2 in neurons could directly influence firing and plasticity by controlling the FGF14 dimerization equilibrium, and thereby the availability of monomeric species for interaction with Nav1.6.
Identifiants
pubmed: 32599005
pii: S0167-4889(20)30144-0
doi: 10.1016/j.bbamcr.2020.118786
pmc: PMC7984254
mid: NIHMS1681046
pii:
doi:
Substances chimiques
NAV1.6 Voltage-Gated Sodium Channel
0
Protein Kinase Inhibitors
0
fibroblast growth factor 14
0
Phosphotyrosine
21820-51-9
Fibroblast Growth Factors
62031-54-3
Janus Kinase 2
EC 2.7.10.2
src-Family Kinases
EC 2.7.10.2
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
118786Subventions
Organisme : NIGMS NIH HHS
ID : T32 GM008280
Pays : United States
Organisme : NIMH NIH HHS
ID : R01 MH124351
Pays : United States
Organisme : NIMH NIH HHS
ID : R01 MH095995
Pays : United States
Organisme : NIH HHS
ID : S10 OD023576
Pays : United States
Organisme : NIMH NIH HHS
ID : R01 MH111107
Pays : United States
Informations de copyright
Copyright © 2020 Elsevier B.V. All rights reserved.
Déclaration de conflit d'intérêts
Declaration of competing interest All authors declare that they have no competing interests.
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