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Purification of Functional Human TRP Channels Recombinantly Produced in Yeast

(1) Background: Human transient receptor potential (TRP) channels constitute a large family of ion-conducting membrane proteins that allow the sensation of environmental cues. As the dysfunction of TRP channels contributes to the pathogenesis of many widespread diseases, including cardiac disorders,...

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Autores principales: Zhang, Liying, Wang, Kaituo, Klaerke, Dan Arne, Calloe, Kirstine, Lowrey, Lillian, Pedersen, Per Amstrup, Gourdon, Pontus, Gotfryd, Kamil
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6406451/
https://www.ncbi.nlm.nih.gov/pubmed/30754715
http://dx.doi.org/10.3390/cells8020148
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author Zhang, Liying
Wang, Kaituo
Klaerke, Dan Arne
Calloe, Kirstine
Lowrey, Lillian
Pedersen, Per Amstrup
Gourdon, Pontus
Gotfryd, Kamil
author_facet Zhang, Liying
Wang, Kaituo
Klaerke, Dan Arne
Calloe, Kirstine
Lowrey, Lillian
Pedersen, Per Amstrup
Gourdon, Pontus
Gotfryd, Kamil
author_sort Zhang, Liying
collection PubMed
description (1) Background: Human transient receptor potential (TRP) channels constitute a large family of ion-conducting membrane proteins that allow the sensation of environmental cues. As the dysfunction of TRP channels contributes to the pathogenesis of many widespread diseases, including cardiac disorders, these proteins also represent important pharmacological targets. TRP channels are typically produced using expensive and laborious mammalian or insect cell-based systems. (2) Methods: We demonstrate an alternative platform exploiting the yeast Saccharomyces cerevisiae capable of delivering high yields of functional human TRP channels. We produce 11 full-length human TRP members originating from four different subfamilies, purify a selected subset of these to a high homogeneity and confirm retained functionality using TRPM8 as a model target. (3) Results: Our findings demonstrate the potential of the described production system for future functional, structural and pharmacological studies of human TRP channels.
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spelling pubmed-64064512019-03-19 Purification of Functional Human TRP Channels Recombinantly Produced in Yeast Zhang, Liying Wang, Kaituo Klaerke, Dan Arne Calloe, Kirstine Lowrey, Lillian Pedersen, Per Amstrup Gourdon, Pontus Gotfryd, Kamil Cells Article (1) Background: Human transient receptor potential (TRP) channels constitute a large family of ion-conducting membrane proteins that allow the sensation of environmental cues. As the dysfunction of TRP channels contributes to the pathogenesis of many widespread diseases, including cardiac disorders, these proteins also represent important pharmacological targets. TRP channels are typically produced using expensive and laborious mammalian or insect cell-based systems. (2) Methods: We demonstrate an alternative platform exploiting the yeast Saccharomyces cerevisiae capable of delivering high yields of functional human TRP channels. We produce 11 full-length human TRP members originating from four different subfamilies, purify a selected subset of these to a high homogeneity and confirm retained functionality using TRPM8 as a model target. (3) Results: Our findings demonstrate the potential of the described production system for future functional, structural and pharmacological studies of human TRP channels. MDPI 2019-02-11 /pmc/articles/PMC6406451/ /pubmed/30754715 http://dx.doi.org/10.3390/cells8020148 Text en © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Zhang, Liying
Wang, Kaituo
Klaerke, Dan Arne
Calloe, Kirstine
Lowrey, Lillian
Pedersen, Per Amstrup
Gourdon, Pontus
Gotfryd, Kamil
Purification of Functional Human TRP Channels Recombinantly Produced in Yeast
title Purification of Functional Human TRP Channels Recombinantly Produced in Yeast
title_full Purification of Functional Human TRP Channels Recombinantly Produced in Yeast
title_fullStr Purification of Functional Human TRP Channels Recombinantly Produced in Yeast
title_full_unstemmed Purification of Functional Human TRP Channels Recombinantly Produced in Yeast
title_short Purification of Functional Human TRP Channels Recombinantly Produced in Yeast
title_sort purification of functional human trp channels recombinantly produced in yeast
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6406451/
https://www.ncbi.nlm.nih.gov/pubmed/30754715
http://dx.doi.org/10.3390/cells8020148
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