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The application of the Escherichia coli giant spheroplast for drug screening with automated planar patch clamp system

Kv2.1, the voltage-gated ion channel, is ubiquitously expressed in variety of tissues and dysfunction of this ion channel is responsible for multiple diseases. Electrophysiological properties of ion channels are so far characterized with eukaryotic cells using the manual patch clamp which requires s...

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Detalles Bibliográficos
Autores principales: Kikuchi, Kyoko, Sugiura, Mika, Nishizawa-Harada, Chizuko, Kimura, Tadashi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5466043/
https://www.ncbi.nlm.nih.gov/pubmed/28626710
http://dx.doi.org/10.1016/j.btre.2015.04.007
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author Kikuchi, Kyoko
Sugiura, Mika
Nishizawa-Harada, Chizuko
Kimura, Tadashi
author_facet Kikuchi, Kyoko
Sugiura, Mika
Nishizawa-Harada, Chizuko
Kimura, Tadashi
author_sort Kikuchi, Kyoko
collection PubMed
description Kv2.1, the voltage-gated ion channel, is ubiquitously expressed in variety of tissues and dysfunction of this ion channel is responsible for multiple diseases. Electrophysiological properties of ion channels are so far characterized with eukaryotic cells using the manual patch clamp which requires skilful operators and expensive equipments. In this research, we created a simple and sensitive drug screen method using bacterial giant spheroplasts and the automated patch clamp which does not require special skills. We expressed a eukaryotic voltage-gated ion channel Kv2.1 in Escherichia coli using prokaryotic codon, and prepared giant spheroplasts large enough for the patch clamp. Human Kv2.1 currents were successfully recorded from giant spheroplasts with the automated system, and Kv2.1-expressed E. coli spheroplasts could steadily reacted to the dose–response assay with TEA and 4-AP. Collectively, our results indicate for the first time that the bacterial giant spheroplast can be applied for practical pharmaceutical assay using the automated patch clamp.
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spelling pubmed-54660432017-06-16 The application of the Escherichia coli giant spheroplast for drug screening with automated planar patch clamp system Kikuchi, Kyoko Sugiura, Mika Nishizawa-Harada, Chizuko Kimura, Tadashi Biotechnol Rep (Amst) Article Kv2.1, the voltage-gated ion channel, is ubiquitously expressed in variety of tissues and dysfunction of this ion channel is responsible for multiple diseases. Electrophysiological properties of ion channels are so far characterized with eukaryotic cells using the manual patch clamp which requires skilful operators and expensive equipments. In this research, we created a simple and sensitive drug screen method using bacterial giant spheroplasts and the automated patch clamp which does not require special skills. We expressed a eukaryotic voltage-gated ion channel Kv2.1 in Escherichia coli using prokaryotic codon, and prepared giant spheroplasts large enough for the patch clamp. Human Kv2.1 currents were successfully recorded from giant spheroplasts with the automated system, and Kv2.1-expressed E. coli spheroplasts could steadily reacted to the dose–response assay with TEA and 4-AP. Collectively, our results indicate for the first time that the bacterial giant spheroplast can be applied for practical pharmaceutical assay using the automated patch clamp. Elsevier 2015-05-05 /pmc/articles/PMC5466043/ /pubmed/28626710 http://dx.doi.org/10.1016/j.btre.2015.04.007 Text en © 2015 The Authors http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Article
Kikuchi, Kyoko
Sugiura, Mika
Nishizawa-Harada, Chizuko
Kimura, Tadashi
The application of the Escherichia coli giant spheroplast for drug screening with automated planar patch clamp system
title The application of the Escherichia coli giant spheroplast for drug screening with automated planar patch clamp system
title_full The application of the Escherichia coli giant spheroplast for drug screening with automated planar patch clamp system
title_fullStr The application of the Escherichia coli giant spheroplast for drug screening with automated planar patch clamp system
title_full_unstemmed The application of the Escherichia coli giant spheroplast for drug screening with automated planar patch clamp system
title_short The application of the Escherichia coli giant spheroplast for drug screening with automated planar patch clamp system
title_sort application of the escherichia coli giant spheroplast for drug screening with automated planar patch clamp system
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5466043/
https://www.ncbi.nlm.nih.gov/pubmed/28626710
http://dx.doi.org/10.1016/j.btre.2015.04.007
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