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

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 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 char

SUBMITTER: Winterstein LM 

PROVIDER: S-EPMC5881443 | biostudies-literature | 2018 Apr

REPOSITORIES: biostudies-literature