Winterstein, Laura-Marie ; Kukovetz, Kerri ; Rauh, Oliver ; Turman, Daniel L. ; Braun, Christian ; Moroni, Anna ; Schroeder, Indra ; Thiel, Gerhard (2018)
Reconstitution and functional characterization of ion channels from nanodiscs in lipid bilayers.
In: The Journal of general physiology, 150 (4)
Artikel, Bibliographie
Dies ist die neueste Version dieses Eintrags.
Kurzbeschreibung (Abstract)
Recent studies have shown that membrane proteins can be efficiently synthesized in vitro before spontaneously inserting into soluble nanoscale lipid bilayers called nanodiscs (NDs). In this paper, we present experimental details that allow a combination of in vitro translation of ion channels into commercially available NDs followed by their direct reconstitution from these nanobilayers into standard bilayer setups for electrophysiological characterization. We present data showing that two model Kchannels, Kcv and KcsA, as well as a recently discovered dual-topology Fchannel, Fluc, can be reliably reconstituted from different types of NDs into bilayers without contamination from the in vitro translation cocktail. The functional properties of Kcv and KcsA were characterized electrophysiologically and exhibited sensitivity to the lipid composition of the target DPhPC bilayer, suggesting that the channel proteins were fully exposed to the target membrane and were no longer surrounded by the lipid/protein scaffold. The single-channel properties of the three tested channels are compatible with studies from recordings of the same proteins in other expression systems. Altogether, the data show that synthesis of ion channels into NDs and their subsequent reconstitution into conventional bilayers provide a fast and reliable method for functional analysis of ion channels.
| Typ des Eintrags: | Artikel |
|---|---|
| Erschienen: | 2018 |
| Autor(en): | Winterstein, Laura-Marie ; Kukovetz, Kerri ; Rauh, Oliver ; Turman, Daniel L. ; Braun, Christian ; Moroni, Anna ; Schroeder, Indra ; Thiel, Gerhard |
| Art des Eintrags: | Bibliographie |
| Titel: | Reconstitution and functional characterization of ion channels from nanodiscs in lipid bilayers. |
| Sprache: | Englisch |
| Publikationsjahr: | April 2018 |
| Titel der Zeitschrift, Zeitung oder Schriftenreihe: | The Journal of general physiology |
| Jahrgang/Volume einer Zeitschrift: | 150 |
| (Heft-)Nummer: | 4 |
| Zugehörige Links: | |
| Kurzbeschreibung (Abstract): | Recent studies have shown that membrane proteins can be efficiently synthesized in vitro before spontaneously inserting into soluble nanoscale lipid bilayers called nanodiscs (NDs). In this paper, we present experimental details that allow a combination of in vitro translation of ion channels into commercially available NDs followed by their direct reconstitution from these nanobilayers into standard bilayer setups for electrophysiological characterization. We present data showing that two model Kchannels, Kcv and KcsA, as well as a recently discovered dual-topology Fchannel, Fluc, can be reliably reconstituted from different types of NDs into bilayers without contamination from the in vitro translation cocktail. The functional properties of Kcv and KcsA were characterized electrophysiologically and exhibited sensitivity to the lipid composition of the target DPhPC bilayer, suggesting that the channel proteins were fully exposed to the target membrane and were no longer surrounded by the lipid/protein scaffold. The single-channel properties of the three tested channels are compatible with studies from recordings of the same proteins in other expression systems. Altogether, the data show that synthesis of ion channels into NDs and their subsequent reconstitution into conventional bilayers provide a fast and reliable method for functional analysis of ion channels. |
| ID-Nummer: | pmid:29487088 |
| 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 Mär 2018 09:28 |
| Letzte Änderung: | 03 Jul 2024 02:28 |
| PPN: | |
| Export: | |
| Suche nach Titel in: | TUfind oder in Google |
Verfügbare Versionen dieses Eintrags
-
Reconstitution and functional characterization of ion channels from nanodiscs in lipid bilayers. (deposited 01 Mär 2022 13:19)
- Reconstitution and functional characterization of ion channels from nanodiscs in lipid bilayers. (deposited 07 Mär 2018 09:28) [Gegenwärtig angezeigt]
 |
Frage zum Eintrag |
Optionen (nur für Redakteure)
 |
Redaktionelle Details anzeigen |
Drucken
Impressum