Inhibition of the Ubc9 E2 SUMO-conjugating enzyme-CRMP2 interaction decreases NaV1.7 currents and reverses experimental neuropathic pain

Liberty François-Moutal, Erik T. Dustrude, Yue Wang, Tatiana Brustovetsky, Angie Dorame, Weina Ju, Aubin Moutal, Samantha Perez-Miller, Nikolai Broustovetski, Vijay Gokhale, May Khanna, Rajesh Khanna

Research output: Contribution to journalArticle

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Abstract

We previously reported that destruction of the small ubiquitin-like modifier (SUMO) modification site in the axonal collapsin response mediator protein 2 (CRMP2) was sufficient to selectively decrease trafficking of the voltage-gated sodium channel NaV1.7 and reverse neuropathic pain. Here, we further interrogate the biophysical nature of the interaction between CRMP2 and the SUMOylation machinery, and test the hypothesis that a rationally designed CRMP2 SUMOylation motif (CSM) peptide can interrupt E2 SUMO-conjugating enzyme Ubc9-dependent modification of CRMP2 leading to a similar suppression of NaV1.7 currents. Microscale thermophoresis and amplified luminescent proximity homogeneous alpha assay revealed a low micromolar binding affinity between CRMP2 and Ubc9. A heptamer peptide harboring CRMP2's SUMO motif, also bound with similar affinity to Ubc9, disrupted the CRMP2-Ubc9 interaction in a concentration-dependent manner. Importantly, incubation of a tat-conjugated cell-penetrating peptide (t-CSM) decreased sodium currents, predominantly NaV1.7, in a model neuronal cell line. Dialysis of t-CSM peptide reduced CRMP2 SUMOylation and blocked surface trafficking of NaV1.7 in rat sensory neurons. Fluorescence dye-based imaging in rat sensory neurons demonstrated inhibition of sodium influx in the presence of t-CSM peptide; by contrast, calcium influx was unaffected. Finally, t-CSM effectively reversed persistent mechanical and thermal hypersensitivity induced by a spinal nerve injury, a model of neuropathic pain. Structural modeling has now identified a pocket-harboring CRMP2's SUMOylation motif that, when targeted through computational screening of ligands/molecules, is expected to identify small molecules that will biochemically and functionally target CRMP2's SUMOylation to reduce NaV1.7 currents and reverse neuropathic pain.

Original languageEnglish (US)
Pages (from-to)2115-2127
Number of pages13
JournalPain
Volume159
Issue number10
DOIs
StatePublished - Oct 1 2018

Fingerprint

Sumoylation
Neuralgia
Ubiquitin
Enzymes
Peptides
Sensory Receptor Cells
NAV1.7 Voltage-Gated Sodium Channel
Sodium
Cell-Penetrating Peptides
collapsin response mediator protein-2
Inhibition (Psychology)
Spinal Injuries
Spinal Nerves
Dialysis
Hypersensitivity
Coloring Agents
Hot Temperature
Fluorescence
Ligands
Calcium

ASJC Scopus subject areas

  • Neurology
  • Clinical Neurology
  • Anesthesiology and Pain Medicine

Cite this

François-Moutal, L., Dustrude, E. T., Wang, Y., Brustovetsky, T., Dorame, A., Ju, W., ... Khanna, R. (2018). Inhibition of the Ubc9 E2 SUMO-conjugating enzyme-CRMP2 interaction decreases NaV1.7 currents and reverses experimental neuropathic pain. Pain, 159(10), 2115-2127. https://doi.org/10.1097/j.pain.0000000000001294

Inhibition of the Ubc9 E2 SUMO-conjugating enzyme-CRMP2 interaction decreases NaV1.7 currents and reverses experimental neuropathic pain. / François-Moutal, Liberty; Dustrude, Erik T.; Wang, Yue; Brustovetsky, Tatiana; Dorame, Angie; Ju, Weina; Moutal, Aubin; Perez-Miller, Samantha; Broustovetski, Nikolai; Gokhale, Vijay; Khanna, May; Khanna, Rajesh.

In: Pain, Vol. 159, No. 10, 01.10.2018, p. 2115-2127.

Research output: Contribution to journalArticle

François-Moutal, L, Dustrude, ET, Wang, Y, Brustovetsky, T, Dorame, A, Ju, W, Moutal, A, Perez-Miller, S, Broustovetski, N, Gokhale, V, Khanna, M & Khanna, R 2018, 'Inhibition of the Ubc9 E2 SUMO-conjugating enzyme-CRMP2 interaction decreases NaV1.7 currents and reverses experimental neuropathic pain', Pain, vol. 159, no. 10, pp. 2115-2127. https://doi.org/10.1097/j.pain.0000000000001294
François-Moutal, Liberty ; Dustrude, Erik T. ; Wang, Yue ; Brustovetsky, Tatiana ; Dorame, Angie ; Ju, Weina ; Moutal, Aubin ; Perez-Miller, Samantha ; Broustovetski, Nikolai ; Gokhale, Vijay ; Khanna, May ; Khanna, Rajesh. / Inhibition of the Ubc9 E2 SUMO-conjugating enzyme-CRMP2 interaction decreases NaV1.7 currents and reverses experimental neuropathic pain. In: Pain. 2018 ; Vol. 159, No. 10. pp. 2115-2127.
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