Mapping protein interactions of sodium channel NaV1.7 using epitope‐tagged gene‐targeted mice |
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| Authors: | Honglei Huang Martina Pyrski Queensta Millet Stéphane Lolignier Toru Morohashi Samuel J Gossage Maude Jay John E Linley Georgios Baskozos Benedikt M Kessler James J Cox Annette C Dolphin Frank Zufall John N Wood Jing Zhao |
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| Affiliation: | 1. TDI Mass Spectrometry Laboratory, Target Discovery Institute, University of Oxford, Oxford, UK;2. Center for Integrative Physiology and Molecular Medicine, Saarland University, Homburg, Germany;3. Molecular Nociception Group, WIBR, University College London, London, UK;4. Université Clermont Auvergne, Inserm U1107 Neuro‐Dol, Pharmacologie Fondamentale et Clinique de la Douleur, Clermont‐Ferrand, France;5. Neuroscience, IMED Biotech Unit, AstraZeneca, Cambridge, UK;6. Division of Bioscience, University College London, London, UK;7. Department of Neuroscience, Physiology and Pharmacology, University College London, London, UK |
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| 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. |
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| Keywords: | NaV1.7 pain protein– protein interactor sensory neuron sodium channel |
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