Reconstitution and Electrophysiological Characterization of Ion Channels in Lipid Bilayers

2018 ◽  
Vol 81 (1) ◽  
pp. e37 ◽  
Author(s):  
Dan A. Klaerke ◽  
Maria de los Angeles Tejada ◽  
Vibeke Grøsfjeld Christensen ◽  
Mette Lassen ◽  
Per Amstrup Pedersen ◽  
...  
Keyword(s):  
Ion Channels ◽  
Lipid Bilayers ◽  
Electrophysiological Characterization

scholarly journals Reconstitution and functional characterization of ion channels from nanodiscs in lipid bilayers

2018 ◽  
Vol 150 (4) ◽  
pp. 637-646 ◽  
Author(s):  
Laura-Marie Winterstein ◽  
Kerri Kukovetz ◽  
Oliver Rauh ◽  
Daniel L. Turman ◽  
Christian Braun ◽  
...  
Keyword(s):  
Ion Channels ◽  
Lipid Bilayers ◽  
Single Channel ◽  
Functional Characterization ◽  
In Vitro Translation ◽  
Target Membrane ◽  
Electrophysiological Characterization ◽  
Channel Properties

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 K+ channels, Kcv and KcsA, as well as a recently discovered dual-topology F− channel, 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.

scholarly journals Electrophysiological Characterization of Mechanosensitive Ion Channels in Native Bacterial Membranes

2013 ◽  
Vol 104 (2) ◽  
pp. 470a
Author(s):  
Maria Barthmes ◽  
Mac Donald F. Jose ◽  
Niels Fertig ◽  
Christian Wahl-Schott ◽  
Armagan Kocer ◽  
...  
Keyword(s):  
Ion Channels ◽  
Bacterial Membranes ◽  
Mechanosensitive Ion Channels ◽  
Electrophysiological Characterization

scholarly journals Electrophysiological Characterization of two Novel Ion Channels of Mitochondria

2016 ◽  
Vol 110 (3) ◽  
pp. 609a
Author(s):  
Vanessa Checchetto ◽  
Angela Paggio ◽  
Simona Reina ◽  
Diego De Stefani ◽  
Vito De Pinto ◽  
...  
Keyword(s):  
Ion Channels ◽  
Electrophysiological Characterization

scholarly journals Molecular and electrophysiological characterization of transient receptor potential ion channels in the primary murine megakaryocyte

2006 ◽  
Vol 576 (1) ◽  
pp. 151-162 ◽  
Author(s):  
Richard N. Carter ◽  
Gwen Tolhurst ◽  
Gemma Walmsley ◽  
Matthieu Vizuete-Forster ◽  
Nigel Miller ◽  
...  
Keyword(s):  
Ion Channels ◽  
Transient Receptor Potential ◽  
Receptor Potential ◽  
Transient Receptor ◽  
Electrophysiological Characterization
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