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Mapping protein interactions of sodium channel NaV1.7 using epitope-tagged gene targeted mice

ABSTRACT: The voltage-gated sodium channel NaV1.7 plays a critical role in pain pathways. As well as action potential propagation, NaV1.7 regulates neurotransmitter release, integrates depolarizing inputs over long periods and regulates transcription. In order to better understand these functions, we generated an epitope-tagged NaV1.7 mouse that showed normal NaV1.7 channel activity and normal pain behavior. Analysis of NaV1.7 complexes affinity-purified under native conditions by mass spectrometry revealed 267 NaV1.7 associated proteins including known and novel interactors such as sodium channel β3 subunit (Scn3b) and collapsin response mediator protein (Crmp2). Selected NaV1.7 protein interactors, such as Crmp2, membrane-trafficking protein synapototagmin-2 (Syt2), G protein-regulated inducer of neurite outgrowth 1 (Gprin1), L-type amino acid transporter 1 (Lat1) and transmembrane P24 trafficking protein 10 (Tmed10) were validated using co-immunoprecipitation and functional assays. The information provided with this physiologically normal epitope-tagged mouse should provide useful insights into the downstream mechanisms associated with NaV1.7 channel function.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Brain, Neuron

DISEASE(S): Pain Agnosia

SUBMITTER: honglei huang

LAB HEAD: Benedikt Kessler

PROVIDER: PXD005155 | Pride | 2018-01-15

REPOSITORIES: Pride

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			Action	DRS
	OTE0214_HL_TEV_TAG.mgf	Mgf
	OTE0214_HL_TEV_TAG.raw	Raw
	OTE0214_HL_TEV_WT.mgf	Mgf
	OTE0214_HL_TEV_WT.raw	Raw
	OTE0231_JingZhao_HOMO_K1_2.mgf	Mgf

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