ArticleProtein expression and purification2015
Cell-free expression of a functional pore-only sodium channel.
Article in Protein expression and purification, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 6 papers.
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Who cites it
6 citing papers in PubMed.
- High-yield production and characterization of functional Kir2.1 ion channel usingSynthetic and systems biotechnology · 2025Article
- Ion channel selectivity through ion-modulated changes of selectivity filter pProceedings of the National Academy of Sciences of the United States of America · 2023Article
- Cell-Free Protein Synthesis: A Promising Option for Future Drug Development.BioDrugs : clinical immunotherapeutics, biopharmaceuticals and gene therapy · 2020Review
- Molecular Dynamics of Ion Conduction through the Selectivity Filter of the NaThe journal of physical chemistry. B · 2018Article
- Reconstitution and functional characterization of ion channels from nanodiscs in lipid bilayers.The Journal of general physiology · 2018Article
- Identification and functional characterization of the Piezo1 channel pore domain.The Journal of biological chemistryArticle
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Authors and funding
6 authors.
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Abstract
Voltage-gated sodium channels participate in the propagation of action potentials in excitable cells. Eukaryotic Navs are pseudo homotetrameric polypeptides, comprising four repeats of six transmembrane segments (S1-S6). The first four segments form the voltage-sensing domain and S5 and S6 create the pore domain with the selectivity filter. Prokaryotic Navs resemble these characteristics, but are truly tetrameric. They can typically be efficiently synthesized in bacteria, but production in vitro with cell-free synthesis has not been demonstrated. Here we report the cell-free expression and purification of a prokaryotic tetrameric pore-only sodium channel. We produced milligram quantities of the functional channel protein as characterized by size-exclusion chromatography, infrared spectroscopy and electrophysiological recordings. Cell-free expression enables advanced site-directed labelling, post-translational modifications, and special solubilization schemes. This enables next-generation biophysical experiments to study the principle of sodium ion selectivity and transport in sodium channels.
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