Lipid reconstitution and recording of recombinant ion channels

Methods Enzymol. 2015:556:385-404. doi: 10.1016/bs.mie.2014.12.028. Epub 2015 Mar 21.

Abstract

Ion channels are membrane proteins that facilitate passive ion movements down the electrochemical gradients across biological membranes. They regulate a wide range of important physiological functions, from neuronal excitation, muscle contraction, substance absorption in epithelial tissues, to innate immunity. In recent years, the membrane transport field has seen tremendous advances in membrane protein purification and structural determination. This paves the way for investigators to reconstitute ion channels into biochemically defined lipid systems free of other biological constituents and to study channel functions within a structural context. The goal of this chapter is to guide readers through the process of producing proteoliposomes, designing liposome assays for initial characterization of channel function, and performing electrical recordings for in-depth mechanistic investigation.

Keywords: Electrical recording; Ion channel; Lipid bilayer; Liposome; Membrane protein; Reconstitution.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Bacteria / metabolism
  • Bacterial Proteins / metabolism
  • Electrophysiological Phenomena
  • Electrophysiology / instrumentation*
  • Equipment Design
  • Humans
  • Ion Channels / metabolism*
  • Lipid Bilayers / metabolism*
  • Liposomes / metabolism*
  • Proteolipids / metabolism
  • Recombinant Proteins / metabolism

Substances

  • Bacterial Proteins
  • Ion Channels
  • Lipid Bilayers
  • Liposomes
  • Proteolipids
  • Recombinant Proteins
  • proteoliposomes