Immobilizing channel molecules in artificial lipid bilayers for simultaneous electrical and optical single channel recordings

Toru Ide, Minako Hirano, Takehiko Ichikawa

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

There has been much interest in imaging single drug bindings to ion channel proteins while simultaneously recording single channel current. We developed an experimental apparatus for simultaneous optical and electrical measurement of single channel proteins by combining the single molecule imaging technique and the artificial bilayer technique. However, one major problem is that single molecule imaging of drug bindings is limited by the innate thermal diffusion of channel proteins in the artificial bilayer. Therefore, immobilizing channel proteins in the bilayers is imperative for stable measurements of channel-drug interactions. For future studies on channel-drug interactions, we describe here three different methods for simultaneous optical and electrical observation of single channels in which channel proteins are immobilized. (i) Membrane binding protein annexin V reduces the lateral diffusion of single channel proteins in a concentration-dependent manner. (ii) Channel proteins are immobilized by anchorage through a polyethylene glycol (PEG) molecule to the glass substrate. (iii) Channels immobilized on a gel bead can be directly incorporated into artificial bilayers.

Original languageEnglish
Title of host publicationCell Signaling Reactions
Subtitle of host publicationSingle-Molecular Kinetic Analysis
PublisherSpringer Netherlands
Pages107-120
Number of pages14
ISBN (Print)9789048198634
DOIs
Publication statusPublished - Dec 1 2011
Externally publishedYes

Keywords

  • Annexin
  • BK-channel
  • Bilayer
  • Channel
  • Co affinity gel
  • Conductance
  • Detergent
  • Diffusion
  • Histidine tag
  • Immobilization
  • KcsA
  • MthK
  • Phospholipid
  • Polyethyleneglycol (PEG)
  • Ryanodine receptor (RyR)
  • Single channel
  • Supported bilayer
  • Total internal reflection fluorescence (TIRF) microscope
  • Vesicle fusion

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

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  • Cite this

    Ide, T., Hirano, M., & Ichikawa, T. (2011). Immobilizing channel molecules in artificial lipid bilayers for simultaneous electrical and optical single channel recordings. In Cell Signaling Reactions: Single-Molecular Kinetic Analysis (pp. 107-120). Springer Netherlands. https://doi.org/10.1007/978-90-481-9864-1_5