Immobilizing BK-channels in artificial lipid bilayers using annexin v

Yuko Takeuchi, Takaaki Aoki, Toshio Yanagida, Toru Ide

Research output: Contribution to journalArticle

Abstract

In this report, we show an improved method for the simultaneous measurement of optical and electrical properties of single-channel proteins for analysis of the gating mechanism. Large-conductance Ca2+-activated potassium (BK) channels were isolated from porcine uterine smooth muscle and labeled with Cy5 via antibody against the N-terminal. These Cy5-labeled BK channels were incorporated into lipid bilayer membranes followed by single channel current measurements. Cy5-labeled BK channels possessed Ca2+ and voltage sensitivity. The orientation of the vesicles was determined to be outside-out. Charybdotoxin applied from the cis side blocked the channel current. For stable observations of ligand and channel binding, BK channel immobilization was also examined. The lateral diffusion coefficient of BK channels decreased over 200 fold in 1 μM annexin V while the open probability did not change. This study is a significant advancement in simultaneous measurements of ligand binding and current change at the single channel level.

Original languageEnglish
Pages (from-to)1-5
Number of pages5
Journale-Journal of Surface Science and Nanotechnology
Volume5
DOIs
Publication statusPublished - Jan 15 2007

Keywords

  • Bioimaging and engineering
  • Biophysics
  • Electrophysiology
  • Immobilization
  • Ion channel
  • Molecular dynamics

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Condensed Matter Physics
  • Mechanics of Materials
  • Surfaces and Interfaces
  • Surfaces, Coatings and Films

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