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Heterologous expression and purification of an active human TRPV3 ion channel.

Kol, Stefan ; Braun, Christian ; Thiel, Gerhard ; Doyle, Declan A. ; Sundström, Michael ; Gourdon, Pontus ; Nissen, Poul (2013)
Heterologous expression and purification of an active human TRPV3 ion channel.
In: The FEBS journal, 280 (23)
Artikel, Bibliographie

Kurzbeschreibung (Abstract)

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 has been reported to date. 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.

Typ des Eintrags: Artikel
Erschienen: 2013
Autor(en): Kol, Stefan ; Braun, Christian ; Thiel, Gerhard ; Doyle, Declan A. ; Sundström, Michael ; Gourdon, Pontus ; Nissen, Poul
Art des Eintrags: Bibliographie
Titel: Heterologous expression and purification of an active human TRPV3 ion channel.
Sprache: Englisch
Publikationsjahr: 2013
Titel der Zeitschrift, Zeitung oder Schriftenreihe: The FEBS journal
Jahrgang/Volume einer Zeitschrift: 280
(Heft-)Nummer: 23
Kurzbeschreibung (Abstract):

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 has been reported to date. 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.

Fachbereich(e)/-gebiet(e): 10 Fachbereich Biologie
10 Fachbereich Biologie > Plant Membrane Biophyscis (am 20.12.23 umbenannt in Biologie der Algen und Protozoen)
Hinterlegungsdatum: 07 Jan 2014 12:49
Letzte Änderung: 07 Jan 2014 12:49
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