Pharmacological inhibition of wee1 kinase selectively modulates the voltage-gated Na+ channel 1.2 macromolecular complex

Nolan M. Dvorak, Cynthia Tapia, Timothy J. Baumgartner, Jully Singh, Fernanda Laezza, Aditya K. Singh

Research output: Contribution to journalArticlepeer-review

3 Scopus citations

Abstract

Voltage-gated Na+ (Nav) channels are a primary molecular determinant of the action potential (AP). Despite the canonical role of the pore-forming α subunit in conferring this function, protein–protein interactions (PPI) between the Nav channel α subunit and its auxiliary proteins are necessary to reconstitute the full physiological activity of the channel and to fine-tune neuronal excitability. In the brain, the Nav channel isoforms 1.2 (Nav 1.2) and 1.6 (Nav 1.6) are enriched, and their activities are differentially regulated by the Nav channel auxiliary protein fibroblast growth factor 14 (FGF14). Despite the known regulation of neuronal Nav channel activity by FGF14, less is known about cellular signaling molecules that might modulate these regulatory effects of FGF14. To that end, and building upon our previous investigations suggesting that neuronal Nav channel activity is regulated by a kinase network involving GSK3, AKT, and Wee1, we interrogate in our current investigation how pharmacological inhibition of Wee1 kinase, a serine/threonine and ty-rosine kinase that is a crucial component of the G2-M cell cycle checkpoint, affects the Nav 1.2 and Nav 1.6 channel macromolecular complexes. Our results show that the highly selective inhibitor of Wee1 kinase, called Wee1 inhibitor II, modulates FGF14:Nav 1.2 complex assembly, but does not significantly affect FGF14:Nav 1.6 complex assembly. These results are functionally recapitulated, as Wee1 inhibitor II entirely alters FGF14-mediated regulation of the Nav 1.2 channel, but displays no effects on the Nav 1.6 channel. At the molecular level, these effects of Wee1 inhibitor II on FGF14:Nav 1.2 complex assembly and FGF14-mediated regulation of Nav 1.2-mediated Na+ currents are shown to be dependent upon the presence of Y158 of FGF14, a residue known to be a prominent site for phosphorylation-mediated regulation of the protein. Overall, our data suggest that pharmacological inhibition of Wee1 confers selective modulatory effects on Nav 1.2 channel activity, which has important implications for unraveling cellular signaling pathways that fine-tune neuronal excitability.

Original languageEnglish (US)
Article number3103
JournalCells
Volume10
Issue number11
DOIs
StatePublished - Nov 2021
Externally publishedYes

Keywords

  • Fibroblast growth factor 14 (FGF14)
  • Patch-clamp electrophysiology
  • Voltage-gated Na (Na) channels
  • Wee1 kinase

ASJC Scopus subject areas

  • General Biochemistry, Genetics and Molecular Biology

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