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Heterodimerization of two pore domain K(+) channel TASK1 and TALK2 in living heterologous expression systems
Two-pore-domain K(+) (K(2P)) channels sense a wide variety of stimuli such as mechanical stress, inhalational anesthetics, and changes in extracellular pH or temperature. The K(2P) channel activity forms a background K(+) current and, thereby, contributes to resting membrane potentials. Six subfamil...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5634629/ https://www.ncbi.nlm.nih.gov/pubmed/29016681 http://dx.doi.org/10.1371/journal.pone.0186252 |
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author | Suzuki, Yoshiaki Tsutsumi, Kanako Miyamoto, Tatsuya Yamamura, Hisao Imaizumi, Yuji |
author_facet | Suzuki, Yoshiaki Tsutsumi, Kanako Miyamoto, Tatsuya Yamamura, Hisao Imaizumi, Yuji |
author_sort | Suzuki, Yoshiaki |
collection | PubMed |
description | Two-pore-domain K(+) (K(2P)) channels sense a wide variety of stimuli such as mechanical stress, inhalational anesthetics, and changes in extracellular pH or temperature. The K(2P) channel activity forms a background K(+) current and, thereby, contributes to resting membrane potentials. Six subfamilies including fifteen subtypes of K(2P) channels have been identified. Each K(2P) channel molecule with two pores consists of a homodimer of each subtype. In addition, a few heterodimers mainly within the same subfamilies have been found recently. In the present study, the possibility of heterodimerization between TASK1 (TWIK-Related Acid-Sensitive K(+) channel) and TALK2 (TWIK-Related Alkaline pH-Activated K(+) channel) was examined. These channels belong to separate subfamilies and show extremely different channel properties. Surprisingly, single molecular imaging analyses in this study using a total internal reflection microscope suggested the heterodimerization of TASK1 and TALK2 in a pancreatic cell line, QGP-1. This heterodimer was also detected using a bimolecular fluorescence complementation assay in a HEK293 heterologous expression system. Fluorescence resonance energy transfer analyses showed that the affinity between TASK1 and TALK2 appeared to be close to those of homodimers. Whole-cell patch-clamp recordings revealed that TASK1 currents in HEK293 cells were significantly attenuated by co-expression of a dominant-negative form of TALK2 in comparison with that of wild-type TALK2. The sensitivities of TASK1-TALK2 tandem constructs to extracellular pH and halothane were characterized as a unique hybrid of TASK1 and TALK2. These results suggested that heterodimerization of TASK1 and TALK2 provides cells with the ability to make multiple responses to a variety of physiological and pharmacological stimuli. |
format | Online Article Text |
id | pubmed-5634629 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-56346292017-10-30 Heterodimerization of two pore domain K(+) channel TASK1 and TALK2 in living heterologous expression systems Suzuki, Yoshiaki Tsutsumi, Kanako Miyamoto, Tatsuya Yamamura, Hisao Imaizumi, Yuji PLoS One Research Article Two-pore-domain K(+) (K(2P)) channels sense a wide variety of stimuli such as mechanical stress, inhalational anesthetics, and changes in extracellular pH or temperature. The K(2P) channel activity forms a background K(+) current and, thereby, contributes to resting membrane potentials. Six subfamilies including fifteen subtypes of K(2P) channels have been identified. Each K(2P) channel molecule with two pores consists of a homodimer of each subtype. In addition, a few heterodimers mainly within the same subfamilies have been found recently. In the present study, the possibility of heterodimerization between TASK1 (TWIK-Related Acid-Sensitive K(+) channel) and TALK2 (TWIK-Related Alkaline pH-Activated K(+) channel) was examined. These channels belong to separate subfamilies and show extremely different channel properties. Surprisingly, single molecular imaging analyses in this study using a total internal reflection microscope suggested the heterodimerization of TASK1 and TALK2 in a pancreatic cell line, QGP-1. This heterodimer was also detected using a bimolecular fluorescence complementation assay in a HEK293 heterologous expression system. Fluorescence resonance energy transfer analyses showed that the affinity between TASK1 and TALK2 appeared to be close to those of homodimers. Whole-cell patch-clamp recordings revealed that TASK1 currents in HEK293 cells were significantly attenuated by co-expression of a dominant-negative form of TALK2 in comparison with that of wild-type TALK2. The sensitivities of TASK1-TALK2 tandem constructs to extracellular pH and halothane were characterized as a unique hybrid of TASK1 and TALK2. These results suggested that heterodimerization of TASK1 and TALK2 provides cells with the ability to make multiple responses to a variety of physiological and pharmacological stimuli. Public Library of Science 2017-10-10 /pmc/articles/PMC5634629/ /pubmed/29016681 http://dx.doi.org/10.1371/journal.pone.0186252 Text en © 2017 Suzuki et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Research Article Suzuki, Yoshiaki Tsutsumi, Kanako Miyamoto, Tatsuya Yamamura, Hisao Imaizumi, Yuji Heterodimerization of two pore domain K(+) channel TASK1 and TALK2 in living heterologous expression systems |
title | Heterodimerization of two pore domain K(+) channel TASK1 and TALK2 in living heterologous expression systems |
title_full | Heterodimerization of two pore domain K(+) channel TASK1 and TALK2 in living heterologous expression systems |
title_fullStr | Heterodimerization of two pore domain K(+) channel TASK1 and TALK2 in living heterologous expression systems |
title_full_unstemmed | Heterodimerization of two pore domain K(+) channel TASK1 and TALK2 in living heterologous expression systems |
title_short | Heterodimerization of two pore domain K(+) channel TASK1 and TALK2 in living heterologous expression systems |
title_sort | heterodimerization of two pore domain k(+) channel task1 and talk2 in living heterologous expression systems |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5634629/ https://www.ncbi.nlm.nih.gov/pubmed/29016681 http://dx.doi.org/10.1371/journal.pone.0186252 |
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