Heterologous expression and purification of an active human TRPV3 ion channel

Publikation: Bidrag til tidsskrift/Konferencebidrag i tidsskrift /Bidrag til avisTidsskriftartikelForskningpeer review

  • Stefan Kol, Protein Production Facility, Danmark
  • Christian Braun, Plant Membrane Biophysics, Technische Universität Darmstadt, Tyskland
  • Gerhard Thiel, Plant Membrane Biophysics, Technische Universität Darmstadt, Tyskland
  • Declan A Doyle, University of Southampton, Storbritannien
  • Michael Sundström, Protein Production Facility, Danmark
  • Pontus Gourdon, Danmark
  • Poul Nissen
The transient receptor potential vanilloid 3 (TRPV3) cation channel is widely expressed in human tissues and has been shown to be activated by mild temperatures or chemical ligands. In spite of great progress in the TRP channel characterization, very little is known about their structure and interactions with other proteins at the atomic level. This is mainly caused by difficulties in obtaining functionally active samples of high homogeneity. Here, we report on the high-level Escherichia coli expression of the human TRPV3 channel, for which no structural information is reported thus far. We selected a suitable detergent and buffer system using analytical size exclusion chromatography and a thermal stability assay. We demonstrate that the recombinant purified protein contains high α-helical content and migrates as dimers and tetramers on native PAGE. Furthermore, the purified channel also retains its current inducing activity as shown by electrophysiology experiments. The ability to produce the TRPV3 channel heterologously will aid future functional and structural studies. This article is protected by copyright. All rights reserved.
OriginalsprogEngelsk
TidsskriftF E B S Journal
Vol/bind280
Nummer23
Sider (fra-til)6010–6021
Antal sider12
ISSN1742-464X
DOI
StatusUdgivet - dec. 2013

Se relationer på Aarhus Universitet Citationsformater

ID: 56336051