(S)-Lacosamide Binding to Collapsin Response Mediator Protein 2 (CRMP2) Regulates CaV2.2 Activity by Subverting Its Phosphorylation by Cdk5

Aubin Moutal, Liberty François-Moutal, Samantha Perez-Miller, Karissa Cottier, Lindsey Anne Chew, Seul Ki Yeon, Jixun Dai, Ki Duk Park, May Khanna, Rajesh Khanna

Research output: Research - peer-reviewArticle

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Abstract

The neuronal circuit remodels during development as well as in human neuropathologies such as epilepsy. Neurite outgrowth is an obligatory step in these events. We recently reported that alterations in the phosphorylation state of an axon specification/guidance protein, the collapsin response mediator protein 2 (CRMP2), play a major role in the activity-dependent regulation of neurite outgrowth. We also identified (S)-LCM, an inactive stereoisomer of the clinically used antiepileptic drug (R)-LCM (Vimpat®), as a novel tool for preferentially targeting CRMP2-mediated neurite outgrowth. Here, we investigated the mechanism by which (S)-LCM affects CRMP2 phosphorylation by two key kinases, cyclin-dependent kinase 5 (Cdk5) and glycogen synthase kinase 3β (GSK-3β). (S)-LCM application to embryonic cortical neurons resulted in reduced levels of Cdk5- and GSK-3β-phosphorylated CRMP2. Mechanistically, (S)-LCM increased CRMP2 binding to both Cdk5- and GSK-3β without affecting binding of CRMP2 to its canonical partner tubulin. Saturation transfer difference nuclear magnetic resonance (STD NMR) and differential scanning fluorimetry (DSF) experiments demonstrated direct binding of (S)-LCM to CRMP2. Using an in vitro luminescent kinase assay, we observed that (S)-LCM specifically inhibited Cdk5-mediated phosphorylation of CRMP2. Cross-linking experiments and analytical ultracentrifugation showed no effect of (S)-LCM on the oligomerization state of CRMP2. The increased association between Cdk5-phosphorylated CRMP2 and CaV2.2 was reduced by (S)-LCM in vitro and in vivo. This reduction translated into a decrease of calcium influx via CaV2.2 in (S)-LCM-treated neurons compared to controls. (S)-LCM, to our knowledge, is the first molecule described to directly inhibit CRMP2 phosphorylation and may be useful for delineating CRMP2-facilitated functions.

LanguageEnglish (US)
Pages1959-1976
Number of pages18
JournalMolecular Neurobiology
Volume53
Issue number3
DOIs
StatePublished - Apr 1 2016

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Cyclin-Dependent Kinase 5
Phosphorylation
collapsin response mediator protein-2
lacosamide
Glycogen Synthase Kinases
Neuronal Outgrowth
Phosphotransferases
Neurons
In Vitro Techniques
Luminescent Measurements
Fluorometry
Stereoisomerism
Ultracentrifugation
Tubulin
Protein Binding
Anticonvulsants
Epilepsy
Magnetic Resonance Spectroscopy
Calcium
Proteins

Keywords

  • (S)-Lacosamide
  • CaV2.2
  • Cdk5
  • CRMP2
  • DSF
  • GSK-3β
  • STD NMR

ASJC Scopus subject areas

  • Cellular and Molecular Neuroscience

Cite this

(S)-Lacosamide Binding to Collapsin Response Mediator Protein 2 (CRMP2) Regulates CaV2.2 Activity by Subverting Its Phosphorylation by Cdk5. / Moutal, Aubin; François-Moutal, Liberty; Perez-Miller, Samantha; Cottier, Karissa; Chew, Lindsey Anne; Yeon, Seul Ki; Dai, Jixun; Park, Ki Duk; Khanna, May; Khanna, Rajesh.

In: Molecular Neurobiology, Vol. 53, No. 3, 01.04.2016, p. 1959-1976.

Research output: Research - peer-reviewArticle

Moutal, A, François-Moutal, L, Perez-Miller, S, Cottier, K, Chew, LA, Yeon, SK, Dai, J, Park, KD, Khanna, M & Khanna, R 2016, '(S)-Lacosamide Binding to Collapsin Response Mediator Protein 2 (CRMP2) Regulates CaV2.2 Activity by Subverting Its Phosphorylation by Cdk5' Molecular Neurobiology, vol 53, no. 3, pp. 1959-1976. DOI: 10.1007/s12035-015-9141-2
Moutal A, François-Moutal L, Perez-Miller S, Cottier K, Chew LA, Yeon SK et al. (S)-Lacosamide Binding to Collapsin Response Mediator Protein 2 (CRMP2) Regulates CaV2.2 Activity by Subverting Its Phosphorylation by Cdk5. Molecular Neurobiology. 2016 Apr 1;53(3):1959-1976. Available from, DOI: 10.1007/s12035-015-9141-2
Moutal, Aubin ; François-Moutal, Liberty ; Perez-Miller, Samantha ; Cottier, Karissa ; Chew, Lindsey Anne ; Yeon, Seul Ki ; Dai, Jixun ; Park, Ki Duk ; Khanna, May ; Khanna, Rajesh. / (S)-Lacosamide Binding to Collapsin Response Mediator Protein 2 (CRMP2) Regulates CaV2.2 Activity by Subverting Its Phosphorylation by Cdk5. In: Molecular Neurobiology. 2016 ; Vol. 53, No. 3. pp. 1959-1976
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