A novel bungarotoxin binding site-tagged construct reveals MAPK-dependent Kv4.2 trafficking
| Autor: | G. Travis Tabor, Jonathan G. Murphy, Jia-Hua Hu, Dax A. Hoffman, Jung M. Park |
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| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
MAPK/ERK pathway Endosome media_common.quotation_subject Hippocampal formation Biology Hippocampus p38 Mitogen-Activated Protein Kinases Article 03 medical and health sciences Cellular and Molecular Neuroscience 0302 clinical medicine Animals Humans Binding site Dipeptidyl-Peptidases and Tripeptidyl-Peptidases Internalization Protein kinase A Protein Kinase Inhibitors Molecular Biology Cells Cultured media_common Mitogen-Activated Protein Kinase Kinases Neurons Binding Sites Kv Channel-Interacting Proteins Cell Biology Transfection Bungarotoxin Bungarotoxins Rats Cell biology Protein Transport HEK293 Cells Shal Potassium Channels 030104 developmental biology cardiovascular system 030217 neurology & neurosurgery Protein Binding |
| Zdroj: | Mol Cell Neurosci |
| ISSN: | 1044-7431 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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