Purification of Functional Human TRP Channels Recombinantly Produced in Yeast

Cells. 2019 Feb 11;8(2):148. doi: 10.3390/cells8020148.

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.

Keywords: Saccharomyces cerevisiae; ion channels; overproduction; production platform; protein purification; sensors; transient receptor potential (TRP) channels; yeast.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Detergents / pharmacology
  • Humans
  • Phylogeny
  • Recombinant Proteins / biosynthesis*
  • Saccharomyces cerevisiae / metabolism*
  • Solubility
  • Transient Receptor Potential Channels / biosynthesis*
  • Transient Receptor Potential Channels / chemistry
  • Transient Receptor Potential Channels / isolation & purification*

Substances

  • Detergents
  • Recombinant Proteins
  • Transient Receptor Potential Channels