The C-Terminal of Na V 1.7 Is Ubiquitinated by NEDD4L.
Katharine M WrightHanjie JiangWendy XiaMichael B MurphyTatiana N BoroninaJustin N NwaforHyoJeon KimAkunna M IheanachoP Aitana AzurmendiRobert N ColePhilip A ColeSandra B GabelliPublished in: ACS bio & med chem Au (2023)
Na V 1.7, the neuronal voltage-gated sodium channel isoform, plays an important role in the human body's ability to feel pain. Mutations within Na V 1.7 have been linked to pain-related syndromes, such as insensitivity to pain. To date, the regulation and internalization mechanisms of the Na V 1.7 channel are not well known at a biochemical level. In this study, we perform biochemical and biophysical analyses that establish that the HECT-type E3 ligase, NEDD4L, ubiquitinates the cytoplasmic C-terminal (CT) region of Na V 1.7. Through in vitro ubiquitination and mass spectrometry experiments, we identify, for the first time, the lysine residues of Na V 1.7 within the CT region that get ubiquitinated. Furthermore, binding studies with an NEDD4L E3 ligase modulator (ubiquitin variant) highlight the dynamic partnership between NEDD4L and Na V 1.7. These investigations provide a framework for understanding how NEDD4L-dependent regulation of the channel can influence the Na V 1.7 function.
Keyphrases
- chronic pain
- mass spectrometry
- pain management
- neuropathic pain
- endothelial cells
- magnetic resonance imaging
- magnetic resonance
- high resolution
- small molecule
- image quality
- contrast enhanced
- brain injury
- dual energy
- liquid chromatography
- positron emission tomography
- cerebral ischemia
- subarachnoid hemorrhage
- high performance liquid chromatography
- dna binding
- case control
- tandem mass spectrometry