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Cells 2019, 8(2), 148; https://doi.org/10.3390/cells8020148

Purification of Functional Human TRP Channels Recombinantly Produced in Yeast

1
Department of Biomedical Sciences, University of Copenhagen, Nørre Allé 14, DK-2200 Copenhagen N, Denmark
2
Department of Veterinary and Animal Sciences, University of Copenhagen, Dyrlægevej 100, DK-1870 Frederiksberg C, Denmark
3
Department of Biology, University of Copenhagen, Universitetsparken 13, DK-2100 Copenhagen OE, Denmark
*
Authors to whom correspondence should be addressed.
Received: 3 January 2019 / Revised: 27 January 2019 / Accepted: 3 February 2019 / Published: 11 February 2019
(This article belongs to the Special Issue TRP Channels in Health and Disease)
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Abstract

(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. View Full-Text
Keywords: ion channels; overproduction; production platform; protein purification; Saccharomyces cerevisiae; sensors; transient receptor potential (TRP) channels; yeast ion channels; overproduction; production platform; protein purification; Saccharomyces cerevisiae; sensors; transient receptor potential (TRP) channels; yeast
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Zhang, L.; Wang, K.; Klaerke, D.A.; Calloe, K.; Lowrey, L.; Pedersen, P.A.; Gourdon, P.; Gotfryd, K. Purification of Functional Human TRP Channels Recombinantly Produced in Yeast. Cells 2019, 8, 148.

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