Chemical shift perturbation mapping of the Ubc9-CRMP2 interface identifies a pocket in CRMP2 amenable for allosteric modulation of Nav1.7 channels.

Abstract

Drug discovery campaigns directly targeting the voltage-gated sodium channel NaV1.7, a highly prized target in chronic pain, have not yet been clinically successful. In a differentiated approach, we demonstrated allosteric control of trafficking and activity of NaV1.7 by prevention of SUMOylation of collapsin response mediator protein 2 (CRMP2). Spinal administration of a SUMOylation incompetent CRMP2 (CRMP2 K374A) significantly attenuated pain behavior in the spared nerve injury (SNI) model of neuropathic pain, underscoring the importance of SUMOylation of CRMP2 as a pathologic event in chronic pain. Using a rational design strategy, we identified a heptamer peptide harboring CRMP2's SUMO motif that disrupted the CRMP2-Ubc9 interaction, inhibited CRMP2 SUMOylation, inhibited NaV1.7 membrane trafficking, and specifically inhibited NaV1.7 sodium influx in sensory neurons. Importantly, this peptide reversed nerve injury-induced thermal and mechanical hypersensitivity in the SNI model, supporting the practicality of discovering pain drugs by indirectly targeting NaV1.7 via prevention of CRMP2 SUMOylation. Here, our goal was to map the unique interface between CRMP2 and Ubc9, the E2 SUMO conjugating enzyme. Using computational and biophysical approaches, we demonstrate the enzyme/substrate nature of Ubc9/CRMP2 binding and identify hot spots on CRMP2 that may form the basis of future drug discovery campaigns disrupting the CRMP2-Ubc9 interaction to recapitulate allosteric regulation of NaV1.7 for pain relief.

• Authors: Liberty François-Moutal, David Donald Scott, Samantha Perez-Miller, Vijay Gokhale, May Khanna, Rajesh Khanna
• Journal: Channels (Austin, Tex.) (Channels (Austin)) Vol. 12 Issue 1 Pg. 219-227 ( 2018) ISSN: 1933-6969 [Electronic] United States
• PMID: 30081699 (Publication Type: Journal Article, Research Support, N.I.H., Extramural)

Chemical References

• Intercellular Signaling Peptides and Proteins
• NAV1.7 Voltage-Gated Sodium Channel
• Nerve Tissue Proteins
• SCN9A protein, human
• collapsin response mediator protein-2
• Ubiquitin-Conjugating Enzymes
• ubiquitin-conjugating enzyme UBC9

Topics

• Allosteric Regulation
• Humans
• Intercellular Signaling Peptides and Proteins (chemistry, genetics, metabolism)
• NAV1.7 Voltage-Gated Sodium Channel (chemistry, metabolism)
• Nerve Tissue Proteins (chemistry, genetics, metabolism)
• Protein Interaction Maps
• Ubiquitin-Conjugating Enzymes (chemistry, genetics, metabolism)