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Changes in ion selectivity following the asymmetrical addition of charge to the selectivity filter of bacterial sodium channels
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Fedorenko, Olena A., Khovanov, Igor A., Roberts, Stephen K. and Guardiani, Carlo (2020) Changes in ion selectivity following the asymmetrical addition of charge to the selectivity filter of bacterial sodium channels. Entropy, 22 (12). 1390. doi:10.3390/e22121390
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Official URL: https://doi.org/10.3390/e22121390
Abstract
Voltage-gated sodium channels (NaVs) play fundamental roles in eukaryotes, but their exceptional size hinders their structural resolution. Bacterial NaVs are simplified homologues of their eukaryotic counterparts, but their use as models of eukaryotic Na+ channels is limited by their homotetrameric structure at odds with the asymmetric Selectivity Filter (SF) of eukaryotic NaVs. This work aims at mimicking the SF of eukaryotic NaVs by engineering radial asymmetry into the SF of bacterial channels. This goal was pursued with two approaches: the co-expression of different monomers of the NaChBac bacterial channel to induce the random assembly of heterotetramers, and the concatenation of four bacterial monomers to form a concatemer that can be targeted by site-specific mutagenesis. Patch-clamp measurements and Molecular Dynamics simulations showed that an additional gating charge in the SF leads to a significant increase in Na+ and a modest increase in the Ca2+ conductance in the NavMs concatemer in agreement with the behavior of the population of random heterotetramers with the highest proportion of channels with charge −5e. We thus showed that charge, despite being important, is not the only determinant of conduction and selectivity, and we created new tools extending the use of bacterial channels as models of eukaryotic counterparts.
| Item Type: | Journal Article | ||||||||||||
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| Subjects: | Q Science > QH Natural history Q Science > QP Physiology |
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| Divisions: | Faculty of Science, Engineering and Medicine > Engineering > Engineering | ||||||||||||
| Library of Congress Subject Headings (LCSH): | Ion channels , Sodium channels , Eukaryotic cells | ||||||||||||
| Journal or Publication Title: | Entropy | ||||||||||||
| Publisher: | M D P I AG | ||||||||||||
| ISSN: | 1099-4300 | ||||||||||||
| Official Date: | 9 December 2020 | ||||||||||||
| Dates: |
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| Volume: | 22 | ||||||||||||
| Number: | 12 | ||||||||||||
| Article Number: | 1390 | ||||||||||||
| DOI: | 10.3390/e22121390 | ||||||||||||
| Status: | Peer Reviewed | ||||||||||||
| Publication Status: | Published | ||||||||||||
| Access rights to Published version: | Open Access | ||||||||||||
| RIOXX Funder/Project Grant: |
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