Phosphorylation of the HCN channel auxiliary subunit TRIP8b is altered in an animal model of temporal lobe epilepsy and modulates channel function

J Biol Chem. 2019 Oct 25;294(43):15743-15758. doi: 10.1074/jbc.RA119.010027. Epub 2019 Sep 5.

Abstract

Temporal lobe epilepsy (TLE) is a prevalent neurological disorder with many patients experiencing poor seizure control with existing anti-epileptic drugs. Thus, novel insights into the mechanisms of epileptogenesis and identification of new drug targets can be transformative. Changes in ion channel function have been shown to play a role in generating the aberrant neuronal activity observed in TLE. Previous work demonstrates that hyperpolarization-activated cyclic nucleotide-gated (HCN) channels regulate neuronal excitability and are mislocalized within CA1 pyramidal cells in a rodent model of TLE. The subcellular distribution of HCN channels is regulated by an auxiliary subunit, tetratricopeptide repeat-containing Rab8b-interacting protein (TRIP8b), and disruption of this interaction correlates with channel mislocalization. However, the molecular mechanisms responsible for HCN channel dysregulation in TLE are unclear. Here we investigated whether changes in TRIP8b phosphorylation are sufficient to alter HCN channel function. We identified a phosphorylation site at residue Ser237 of TRIP8b that enhances binding to HCN channels and influences channel gating by altering the affinity of TRIP8b for the HCN cytoplasmic domain. Using a phosphospecific antibody, we demonstrate that TRIP8b phosphorylated at Ser237 is enriched in CA1 distal dendrites and that phosphorylation is reduced in the kainic acid model of TLE. Overall, our findings indicate that the TRIP8b-HCN interaction can be modulated by changes in phosphorylation and suggest that loss of TRIP8b phosphorylation may affect HCN channel properties during epileptogenesis. These results highlight the potential of drugs targeting posttranslational modifications to restore TRIP8b phosphorylation to reduce excitability in TLE.

Keywords: Ca2+/calmodulin-dependent protein kinase II (CaMKII); HCN; TLE; TRIP8b; channelopathy; epilepsy; ion channel; neuronal excitability; phosphorylation; protein–protein interaction.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Brain / metabolism
  • Calcium-Calmodulin-Dependent Protein Kinase Type 2 / metabolism
  • Dendrites / metabolism
  • Disease Models, Animal
  • Epilepsy, Temporal Lobe / metabolism*
  • Female
  • HEK293 Cells
  • Humans
  • Ion Channel Gating
  • Kainic Acid
  • Membrane Proteins / chemistry
  • Membrane Proteins / metabolism*
  • Mice, Inbred C57BL
  • Peroxins / chemistry
  • Peroxins / metabolism*
  • Phosphorylation
  • Phosphoserine / metabolism
  • Protein Subunits / chemistry
  • Protein Subunits / metabolism*
  • Rats, Sprague-Dawley
  • Reproducibility of Results

Substances

  • Membrane Proteins
  • Peroxins
  • Pex5l protein, mouse
  • Pex5l protein, rat
  • Protein Subunits
  • Phosphoserine
  • Calcium-Calmodulin-Dependent Protein Kinase Type 2
  • Kainic Acid