A novel bungarotoxin binding site-tagged construct reveals MAPK-dependent Kv4.2 trafficking.

G Travis Tabor, Jung M Park, Jonathan G Murphy, Jia-Hua Hu, Dax A Hoffman

Molecular and cellular neurosciences 2019 Jul

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Kv4.2 voltage-gated K+ channel subunits, the primary source of the somatodendritic A-type K+ current in CA1 pyramidal neurons of the hippocampus, play important roles in regulating dendritic excitability and plasticity. To better study the trafficking and subcellular distribution of Kv4.2, we created and characterized a novel Kv4.2 construct encoding a bungarotoxin binding site in the extracellular S3-S4 linker region of the α-subunit. When expressed, this construct can be visualized in living cells after staining with rhodamine-conjugated bungarotoxin. We validated the utility of this construct by visualizing the spontaneous internalization and insertion of Kv4.2 in HEK 293T cells. We further report that Kv4.2 colocalized with several endosome markers in HEK 293T cells. In addition, Kv4.2 internalization is significantly impaired by mitogen-activated protein kinase (MAPK) inhibitors in transfected primary hippocampal neurons. Therefore, this newly developed BBS-Kv4.2 construct provides a novel and powerful tool for studying surface Kv4.2 channel localization and trafficking. Published by Elsevier Inc.

Citation

G Travis Tabor, Jung M Park, Jonathan G Murphy, Jia-Hua Hu, Dax A Hoffman. A novel bungarotoxin binding site-tagged construct reveals MAPK-dependent Kv4.2 trafficking. Molecular and cellular neurosciences. 2019 Jul;98:121-130