Quantitative proteomics reveals protein-protein interactions with fibroblast growth factor 12 as a component of the voltage-gated sodium channel 1.2 (nav1.2) macromolecular complex in Mammalian brain.

Molecular & Cellular Proteomics : MCP
Norelle C WildburgerFernanda Laezza

Abstract

Voltage-gated sodium channels (Nav1.1-Nav1.9) are responsible for the initiation and propagation of action potentials in neurons, controlling firing patterns, synaptic transmission and plasticity of the brain circuit. Yet, it is the protein-protein interactions of the macromolecular complex that exert diverse modulatory actions on the channel, dictating its ultimate functional outcome. Despite the fundamental role of Nav channels in the brain, information on its proteome is still lacking. Here we used affinity purification from crude membrane extracts of whole brain followed by quantitative high-resolution mass spectrometry to resolve the identity of Nav1.2 protein interactors. Of the identified putative protein interactors, fibroblast growth factor 12 (FGF12), a member of the nonsecreted intracellular FGF family, exhibited 30-fold enrichment in Nav1.2 purifications compared with other identified proteins. Using confocal microscopy, we visualized native FGF12 in the brain tissue and confirmed that FGF12 forms a complex with Nav1.2 channels at the axonal initial segment, the subcellular specialized domain of neurons required for action potential initiation. Co-immunoprecipitation studies in a heterologous expression system valid...Continue Reading

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Citations

Feb 26, 2016·Frontiers in Cellular Neuroscience·Musaad A AlshammariFernanda Laezza
Jan 18, 2018·The EMBO Journal·Alexandros H KanellopoulosJing Zhao
Jul 3, 2020·The Journal of Biological Chemistry·Agnes S ZyburaAndy Hudmon
May 5, 2017·Frontiers in Cellular Neuroscience·Jessica Di ReFernanda Laezza
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Aug 27, 2017·Scientific Reports·Norelle C WildburgerDavid L Brody
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Jun 28, 2016·Journal of Proteome Research·Pratibha NarayananManuela Schmidt
Jan 24, 2018·ACS Chemical Neuroscience·Syed R AliFernanda Laezza

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Datasets Mentioned

BETA
PXD000719

Methods Mentioned

BETA
affinity purification
confocal microscopy
co-immunoprecipitation
immunoprecipitation
co-IP

Software Mentioned

Excel
pCLAMP
PEAKS
AlphaView
Tandem
MASCOT
SAS
PEAKS inChorus
Origin
! Tandem

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