A membrane-delimited N-myristoylated CRMP2 peptide aptamer inhibits CaV2.2 trafficking and reverses inflammatory and postoperative pain behaviors

Liberty François-Moutal, Yue Wang, Aubin Moutal, Karissa E. Cottier, Ohannes K. Melemedjian, Xiaofang Yang, Yuying Wang, Weina Ju, Tally M. Largent-Milnes, May Khanna, Todd W Vanderah, Rajesh Khanna

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

Targeting proteins within the N-type voltage-gated calcium channel (CaV2.2) complex has proven to be an effective strategy for developing novel pain therapeutics. We describe a novel peptide aptamer derived from the collapsin response mediator protein 2 (CRMP2), a CaV2.2-regulatory protein. Addition of a 14-carbon myristate group to the peptide (myr-tat-CBD3) tethered it to the membrane of primary sensory neurons near surface CaV2.2. Pull-down studies demonstrated that myr-tat-CBD3 peptide interfered with the CRMP2-CaV2.2 interaction. Quantitative confocal immunofluorescence revealed a pronounced reduction of CaV2.2 trafficking after myr-tat-CBD3 treatment and increased efficiency in disrupting CRMP2-CaV2.2 colocalization compared with peptide tat-CBD3. Consequently, myr-tat-CBD3 inhibited depolarization-induced calcium influx in sensory neurons. Voltage clamp electrophysiology experiments revealed a reduction of Ca2+, but not Na+, currents in sensory neurons after myr-tat-CBD3 exposure. Current clamp electrophysiology experiments demonstrated a reduction in excitability of small-diameter dorsal root ganglion neurons after exposure to myr-tat-CBD3. Myr-tat-CBD3 was effective in significantly attenuating carrageenan-induced thermal hypersensitivity and reversing thermal hypersensitivity induced by a surgical incision of the plantar surface of the rat hind paw, a model of postoperative pain. These effects are compared with those of tat-CBD3-the nonmyristoylated tat-conjugated CRMP2 peptide as well as scrambled versions of CBD3 and CBD3-lacking control peptides. Our results demonstrate that the myristoyl tag enhances intracellular delivery and local concentration of the CRMP2 peptide aptamer near membrane-delimited calcium channels resulting in pronounced interference with the calcium channel complex, superior suppression of calcium influx, and better antinociceptive potential.

Original languageEnglish (US)
Pages (from-to)1247-1264
Number of pages18
JournalPain
Volume156
Issue number7
DOIs
StatePublished - Jul 1 2015

Fingerprint

Peptide Aptamers
Postoperative Pain
Sensory Receptor Cells
Calcium Channels
Membranes
Electrophysiology
Peptides
Hypersensitivity
Hot Temperature
Calcium
Carrageenan
Myristic Acid
Spinal Ganglia
Protein Transport
Fluorescent Antibody Technique
Carbon
collapsin response mediator protein-2
Neurons
Pain
Therapeutics

Keywords

  • Calcium imaging
  • CaV2.2
  • Conditioned place preference
  • CRMP2
  • GPMVs
  • Inflammatory pain
  • N-myristoylated peptide
  • Postoperative pain
  • Trafficking

ASJC Scopus subject areas

  • Clinical Neurology
  • Anesthesiology and Pain Medicine
  • Neurology
  • Pharmacology

Cite this

A membrane-delimited N-myristoylated CRMP2 peptide aptamer inhibits CaV2.2 trafficking and reverses inflammatory and postoperative pain behaviors. / François-Moutal, Liberty; Wang, Yue; Moutal, Aubin; Cottier, Karissa E.; Melemedjian, Ohannes K.; Yang, Xiaofang; Wang, Yuying; Ju, Weina; Largent-Milnes, Tally M.; Khanna, May; Vanderah, Todd W; Khanna, Rajesh.

In: Pain, Vol. 156, No. 7, 01.07.2015, p. 1247-1264.

Research output: Contribution to journalArticle

François-Moutal, L, Wang, Y, Moutal, A, Cottier, KE, Melemedjian, OK, Yang, X, Wang, Y, Ju, W, Largent-Milnes, TM, Khanna, M, Vanderah, TW & Khanna, R 2015, 'A membrane-delimited N-myristoylated CRMP2 peptide aptamer inhibits CaV2.2 trafficking and reverses inflammatory and postoperative pain behaviors', Pain, vol. 156, no. 7, pp. 1247-1264. https://doi.org/10.1097/j.pain.0000000000000147
François-Moutal, Liberty ; Wang, Yue ; Moutal, Aubin ; Cottier, Karissa E. ; Melemedjian, Ohannes K. ; Yang, Xiaofang ; Wang, Yuying ; Ju, Weina ; Largent-Milnes, Tally M. ; Khanna, May ; Vanderah, Todd W ; Khanna, Rajesh. / A membrane-delimited N-myristoylated CRMP2 peptide aptamer inhibits CaV2.2 trafficking and reverses inflammatory and postoperative pain behaviors. In: Pain. 2015 ; Vol. 156, No. 7. pp. 1247-1264.
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