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Development of an automated system to measure ion channel currents using a surface-modified gold probe

Artificial lipid bilayer single-channel recording technique has been employed to determine the biophysical and pharmacological properties of various ion channels. However, its measurement efficiency is very low, as it requires two time-consuming processes: preparation of lipid bilayer membranes and...

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Autores principales: Hirano, Minako, Tomita, Masahisa, Takahashi, Chikako, Kawashima, Nobuyuki, Ide, Toru
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8429628/
https://www.ncbi.nlm.nih.gov/pubmed/34504175
http://dx.doi.org/10.1038/s41598-021-97237-z
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author Hirano, Minako
Tomita, Masahisa
Takahashi, Chikako
Kawashima, Nobuyuki
Ide, Toru
author_facet Hirano, Minako
Tomita, Masahisa
Takahashi, Chikako
Kawashima, Nobuyuki
Ide, Toru
author_sort Hirano, Minako
collection PubMed
description Artificial lipid bilayer single-channel recording technique has been employed to determine the biophysical and pharmacological properties of various ion channels. However, its measurement efficiency is very low, as it requires two time-consuming processes: preparation of lipid bilayer membranes and incorporation of ion channels into the membranes. In order to address these problems, we previously developed a technique based on hydrophilically modified gold probes on which are immobilized ion channels that can be promptly incorporated into the bilayer membrane at the same time as the membrane is formed on the probes’ hydrophilic area. Here, we improved further this technique by optimizing the gold probe and developed an automated channel current measurement system. We found that use of probes with rounded tips enhanced the efficiency of channel current measurements, and introducing a hydrophobic area on the probe surface, beside the hydrophilic one, further increased measurement efficiency by boosting membrane stability. Moreover, we developed an automated measurement system using the optimized probes; it enabled us to automatically measure channel currents and analyze the effects of a blocker on channel activity. Our study will contribute to the development of high-throughput devices to identify drug candidates affecting ion channel activity.
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spelling pubmed-84296282021-09-10 Development of an automated system to measure ion channel currents using a surface-modified gold probe Hirano, Minako Tomita, Masahisa Takahashi, Chikako Kawashima, Nobuyuki Ide, Toru Sci Rep Article Artificial lipid bilayer single-channel recording technique has been employed to determine the biophysical and pharmacological properties of various ion channels. However, its measurement efficiency is very low, as it requires two time-consuming processes: preparation of lipid bilayer membranes and incorporation of ion channels into the membranes. In order to address these problems, we previously developed a technique based on hydrophilically modified gold probes on which are immobilized ion channels that can be promptly incorporated into the bilayer membrane at the same time as the membrane is formed on the probes’ hydrophilic area. Here, we improved further this technique by optimizing the gold probe and developed an automated channel current measurement system. We found that use of probes with rounded tips enhanced the efficiency of channel current measurements, and introducing a hydrophobic area on the probe surface, beside the hydrophilic one, further increased measurement efficiency by boosting membrane stability. Moreover, we developed an automated measurement system using the optimized probes; it enabled us to automatically measure channel currents and analyze the effects of a blocker on channel activity. Our study will contribute to the development of high-throughput devices to identify drug candidates affecting ion channel activity. Nature Publishing Group UK 2021-09-09 /pmc/articles/PMC8429628/ /pubmed/34504175 http://dx.doi.org/10.1038/s41598-021-97237-z Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Hirano, Minako
Tomita, Masahisa
Takahashi, Chikako
Kawashima, Nobuyuki
Ide, Toru
Development of an automated system to measure ion channel currents using a surface-modified gold probe
title Development of an automated system to measure ion channel currents using a surface-modified gold probe
title_full Development of an automated system to measure ion channel currents using a surface-modified gold probe
title_fullStr Development of an automated system to measure ion channel currents using a surface-modified gold probe
title_full_unstemmed Development of an automated system to measure ion channel currents using a surface-modified gold probe
title_short Development of an automated system to measure ion channel currents using a surface-modified gold probe
title_sort development of an automated system to measure ion channel currents using a surface-modified gold probe
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8429628/
https://www.ncbi.nlm.nih.gov/pubmed/34504175
http://dx.doi.org/10.1038/s41598-021-97237-z
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