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

EMBO J. 2018 Feb 1;37(3):427-445. doi: 10.15252/embj.201796692. Epub 2018 Jan 15.

Abstract

The voltage-gated sodium channel NaV1.7 plays a critical role in pain pathways. We generated an epitope-tagged NaV1.7 mouse that showed normal pain behaviours to identify channel-interacting proteins. Analysis of NaV1.7 complexes affinity-purified under native conditions by mass spectrometry revealed 267 proteins associated with Nav1.7 in vivo The sodium channel β3 (Scn3b), rather than the β1 subunit, complexes with Nav1.7, and we demonstrate an interaction between collapsing-response mediator protein (Crmp2) and Nav1.7, through which the analgesic drug lacosamide regulates Nav1.7 current density. Novel NaV1.7 protein interactors including membrane-trafficking protein synaptotagmin-2 (Syt2), L-type amino acid transporter 1 (Lat1) and transmembrane P24-trafficking protein 10 (Tmed10) together with Scn3b and Crmp2 were validated by co-immunoprecipitation (Co-IP) from sensory neuron extract. Nav1.7, known to regulate opioid receptor efficacy, interacts with the G protein-regulated inducer of neurite outgrowth (Gprin1), an opioid receptor-binding protein, demonstrating a physical and functional link between Nav1.7 and opioid signalling. Further information on physiological interactions provided with this normal epitope-tagged mouse should provide useful insights into the many functions now associated with the NaV1.7 channel.

Keywords: NaV1.7; pain; protein–protein interactor; sensory neuron; sodium channel.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetamides / pharmacology
  • Analgesics / pharmacology
  • Animals
  • Cell Line
  • HEK293 Cells
  • Humans
  • Intercellular Signaling Peptides and Proteins / metabolism
  • Lacosamide
  • Mice
  • Mice, Inbred C57BL
  • Mice, Transgenic
  • NAV1.7 Voltage-Gated Sodium Channel / genetics
  • NAV1.7 Voltage-Gated Sodium Channel / metabolism*
  • Nerve Tissue Proteins / metabolism*
  • Pain / physiopathology*
  • Protein Binding
  • Protein Interaction Mapping
  • Protein Transport / physiology
  • Receptors, N-Methyl-D-Aspartate / metabolism*
  • Receptors, Opioid / metabolism*
  • Sensory Receptor Cells / metabolism*
  • Synaptotagmin II / metabolism
  • Vesicular Transport Proteins / metabolism
  • Voltage-Gated Sodium Channel beta-3 Subunit / metabolism

Substances

  • Acetamides
  • Analgesics
  • Gprin1 protein, mouse
  • Intercellular Signaling Peptides and Proteins
  • NAV1.7 Voltage-Gated Sodium Channel
  • Nerve Tissue Proteins
  • Receptors, N-Methyl-D-Aspartate
  • Receptors, Opioid
  • SCN3B protein, mouse
  • Scn9a protein, mouse
  • Synaptotagmin II
  • Syt2 protein, mouse
  • Tmed10 protein, mouse
  • Vesicular Transport Proteins
  • Voltage-Gated Sodium Channel beta-3 Subunit
  • collapsin response mediator protein-2
  • Lacosamide