TY - JOUR
T1 - Dissecting the role of the CRMP2-neurofibromin complex on pain behaviors
AU - Moutal, Aubin
AU - Wang, Yue
AU - Yang, Xiaofang
AU - Ji, Yingshi
AU - Luo, Shizhen
AU - Dorame, Angie
AU - Bellampalli, Shreya S.
AU - Chew, Lindsey A.
AU - Cai, Song
AU - Dustrude, Erik T.
AU - Keener, James E.
AU - Marty, Michael T.
AU - Vanderah, Todd W.
AU - Khanna, Rajesh
N1 - Funding Information:
The authors thank Dr Stephen Sligar (University of Illinois at Urbana-Champaign, IL) for the MSP1E3D1 plasmid and Dr Samantha Perez-Miller for rendering the CRMP2 peptide structure in Pymol. This work was supported by a Neurofibromatosis New Investigator Award from the Department of Defense Congressionally Directed Military Medical Research and Development Program (NF1000099), National Institutes of Health awards (1R01NS098772 and 1R01DA042852), and a Children’s Tumor Foundation NF1 Synodos award (2015-04-009A) to R. Khanna. A. Moutal was supported by a Young Investigator’s Award from the Children’s Tumor Foundation. A. Dorame was supported by funds from the University of Arizona’s Border Latino and American Indian Summer Exposure to Research (BLAISER) program. S. S. Bellampalli and L. A. Chew were supported by funds to the University of Arizona’s Undergraduate Biology Research Program from the
Publisher Copyright:
© 2017 International Association for the Study of Pain.
PY - 2017/11/1
Y1 - 2017/11/1
N2 - Neurofibromatosis type 1 (NF1), a genetic disorder linked to inactivating mutations or a homozygous deletion of the Nf1 gene, is characterized by tumorigenesis, cognitive dysfunction, seizures, migraine, and pain. Omic studies on human NF1 tissues identified an increase in the expression of collapsin response mediator protein 2 (CRMP2), a cytosolic protein reported to regulate the trafficking and activity of presynaptic N-type voltage-gated calcium (Cav2.2) channels. Because neurofibromin, the protein product of the Nf1 gene, binds to and inhibits CRMP2, the neurofibromin-CRMP2 signaling cascade will likely affect Ca 2+ channel activity and regulate nociceptive neurotransmission and in vivo responses to noxious stimulation. Here, we investigated the function of neurofibromin-CRMP2 interaction on Cav2.2. Mapping of >275 peptides between neurofibromin and CRMP2 identified a 15-amino acid CRMP2-derived peptide that, when fused to the tat transduction domain of HIV-1, inhibited Ca 2+ influx in dorsal root ganglion neurons. This peptide mimics the negative regulation of CRMP2 activity by neurofibromin. Neurons treated with tat-CRMP2/neurofibromin regulating peptide 1 (t-CNRP1) exhibited a decreased Cav2.2 membrane localization, and uncoupling of neurofibromin-CRMP2 and CRMP2-Cav2.2 interactions. Proteomic analysis of a nanodisc-solubilized membrane protein library identified syntaxin 1A as a novel CRMP2-binding protein whose interaction with CRMP2 was strengthened in neurofibromin-depleted cells and reduced by t-CNRP1. Stimulus-evoked release of calcitonin gene-related peptide from lumbar spinal cord slices was inhibited by t-CNRP1. Intrathecal administration of t-CNRP1 was antinociceptive in experimental models of inflammatory, postsurgical, and neuropathic pain. Our results demonstrate the utility of t-CNRP1 to inhibit CRMP2 protein-protein interactions for the potential treatment of pain.
AB - Neurofibromatosis type 1 (NF1), a genetic disorder linked to inactivating mutations or a homozygous deletion of the Nf1 gene, is characterized by tumorigenesis, cognitive dysfunction, seizures, migraine, and pain. Omic studies on human NF1 tissues identified an increase in the expression of collapsin response mediator protein 2 (CRMP2), a cytosolic protein reported to regulate the trafficking and activity of presynaptic N-type voltage-gated calcium (Cav2.2) channels. Because neurofibromin, the protein product of the Nf1 gene, binds to and inhibits CRMP2, the neurofibromin-CRMP2 signaling cascade will likely affect Ca 2+ channel activity and regulate nociceptive neurotransmission and in vivo responses to noxious stimulation. Here, we investigated the function of neurofibromin-CRMP2 interaction on Cav2.2. Mapping of >275 peptides between neurofibromin and CRMP2 identified a 15-amino acid CRMP2-derived peptide that, when fused to the tat transduction domain of HIV-1, inhibited Ca 2+ influx in dorsal root ganglion neurons. This peptide mimics the negative regulation of CRMP2 activity by neurofibromin. Neurons treated with tat-CRMP2/neurofibromin regulating peptide 1 (t-CNRP1) exhibited a decreased Cav2.2 membrane localization, and uncoupling of neurofibromin-CRMP2 and CRMP2-Cav2.2 interactions. Proteomic analysis of a nanodisc-solubilized membrane protein library identified syntaxin 1A as a novel CRMP2-binding protein whose interaction with CRMP2 was strengthened in neurofibromin-depleted cells and reduced by t-CNRP1. Stimulus-evoked release of calcitonin gene-related peptide from lumbar spinal cord slices was inhibited by t-CNRP1. Intrathecal administration of t-CNRP1 was antinociceptive in experimental models of inflammatory, postsurgical, and neuropathic pain. Our results demonstrate the utility of t-CNRP1 to inhibit CRMP2 protein-protein interactions for the potential treatment of pain.
KW - Allodynia
KW - CGRP
KW - CRMP2
KW - gp120-induced peripheral neuropathy
KW - Hyperalgesia
KW - Inflammatory pain
KW - N-type voltage-gated calcium channel
KW - Neurofibromatosis type 1
KW - Postsurgical pain
KW - Protein-protein interaction
KW - Proteomics
KW - Syntaxin 1A
KW - Synaptosomes/metabolism
KW - Sensory Receptor Cells/physiology
KW - Male
KW - Intercellular Signaling Peptides and Proteins/metabolism
KW - Hyperalgesia/physiopathology
KW - Pain/etiology
KW - Ganglia, Spinal/pathology
KW - Nerve Tissue Proteins/metabolism
KW - Multiprotein Complexes/metabolism
KW - Reaction Time/drug effects
KW - Female
KW - Disease Models, Animal
KW - Action Potentials/drug effects
KW - Neurofibromin 1/metabolism
KW - Rats
KW - Spinal Cord/metabolism
KW - Ligation/adverse effects
KW - Random Allocation
KW - Rats, Sprague-Dawley
KW - Animals
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U2 - 10.1097/j.pain.0000000000001026
DO - 10.1097/j.pain.0000000000001026
M3 - Article
C2 - 28767512
AN - SCOPUS:85040778191
SN - 0304-3959
VL - 158
SP - 2203
EP - 2221
JO - Pain
JF - Pain
IS - 11
ER -