Molecular Nociception Group, WIBR, University College London, London, UK.
TDI Mass Spectrometry Laboratory, Target Discovery Institute, University of Oxford, Oxford, UK.
EMBO J. 2018 Feb 1;37(3):427-445. doi: 10.15252/embj.201796692. Epub 2018 Jan 15.
The voltage-gated sodium channel Na1.7 plays a critical role in pain pathways. We generated an epitope-tagged Na1.7 mouse that showed normal pain behaviours to identify channel-interacting proteins. Analysis of Na1.7 complexes affinity-purified under native conditions by mass spectrometry revealed 267 proteins associated with Nav1.7 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 Na1.7 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 Na1.7 channel.
电压门控钠离子通道 Na1.7 在疼痛通路中起着关键作用。我们生成了一种标记的 Na1.7 小鼠，该小鼠表现出正常的疼痛行为，以鉴定与通道相互作用的蛋白质。通过质谱分析在天然条件下亲和纯化的 Na1.7 复合物，发现与 Nav1.7 相关的 267 种蛋白质。钠离子通道 β3（Scn3b）而不是 β1 亚基与 Nav1.7 形成复合物，我们证明了 collapsin 反应介质蛋白 2（Crmp2）与 Nav1.7 之间的相互作用，通过这种相互作用，镇痛药物拉科酰胺调节 Nav1.7 电流密度。新型 Na1.7 蛋白相互作用物包括膜转运蛋白突触结合蛋白-2（Syt2）、L 型氨基酸转运蛋白 1（Lat1）和跨膜 P24 转运蛋白 10（Tmed10）与 Scn3b 和 Crmp2 一起通过感觉神经元提取物中的共免疫沉淀（Co-IP）进行验证。已知调节阿片受体效能的 Nav1.7 与 G 蛋白调节的神经突生长诱导物（Gprin1）相互作用，Gprin1 是一种阿片受体结合蛋白，证明了 Nav1.7 与阿片信号之间的物理和功能联系。这种正常标记的小鼠提供的有关生理相互作用的更多信息应该为与 Na1.7 通道相关的许多功能提供有用的见解。
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