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Immobilizing Channel Molecules in Artificial Lipid Bilayers for Simultaneous Electrical and Optical Single Channel Recordings

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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.

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Authors and Affiliations

  1. Graduate School of Frontier Biosciences, Osaka University, 1-3 Yamadaoka Suita, Osaka, 565-0871, Japan

    Toru Ide & Minako Hirano

  2. Laboratory for Spatiotemporal Regulations, National Institute for Basic Biology, Nishigonaka 38, Myodaiji, Okazaki, Aichi, 444-8585, Japan

    Takehiko Ichikawa

Authors
  1. Toru Ide
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  2. Minako Hirano
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  3. Takehiko Ichikawa
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Corresponding author

Correspondence to Toru Ide .

Editor information

Editors and Affiliations

  1. , Cellular Informatics Laboratory, RIKEN, Hirosawa 2-1, Wako, 351-0198, Japan

    Yasushi Sako

  2. Graduate School of Frontier Biosciences, Laboratories of Nanobiology, Osaka University, Yamadaoka 1-3, Suita, 565-871, Japan

    Masahiro Ueda

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Ide, T., Hirano, M., Ichikawa, T. (2011). Immobilizing Channel Molecules in Artificial Lipid Bilayers for Simultaneous Electrical and Optical Single Channel Recordings. In: Sako, Y., Ueda, M. (eds) Cell Signaling Reactions. Springer, Dordrecht. https://doi.org/10.1007/978-90-481-9864-1_5

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