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Arrayed lipid bilayer chambers allow single-molecule analysis of membrane transporter activity

Author

Listed:
  • Rikiya Watanabe

    (Graduate School of Engineering, The University of Tokyo
    PRESTO, JST
    CREST, JST)

  • Naoki Soga

    (Graduate School of Engineering, The University of Tokyo)

  • Daishi Fujita

    (Graduate School of Engineering, The University of Tokyo)

  • Kazuhito V. Tabata

    (Graduate School of Engineering, The University of Tokyo
    PRESTO, JST
    CREST, JST)

  • Lisa Yamauchi

    (Graduate School of Engineering, The University of Tokyo
    CREST, JST)

  • Soo Hyeon Kim

    (CREST, JST
    Institute of Industrial Science, The University of Tokyo)

  • Daisuke Asanuma

    (Laboratory of Neurobiology, Graduate School of Medicine, The University of Tokyo)

  • Mako Kamiya

    (Laboratory of Chemical Biology and Molecular Imaging, Graduate School of Medicine, The University of Tokyo)

  • Yasuteru Urano

    (Laboratory of Chemical Biology and Molecular Imaging, Graduate School of Medicine, The University of Tokyo)

  • Hiroaki Suga

    (CREST, JST
    Graduate School of Science, The University of Tokyo)

  • Hiroyuki Noji

    (Graduate School of Engineering, The University of Tokyo
    CREST, JST)

Abstract

Nano- to micron-size reaction chamber arrays (femtolitre chamber arrays) have facilitated the development of sensitive and quantitative biological assays, such as single-molecule enzymatic assays, digital PCR and digital ELISA. However, the versatility of femtolitre chamber arrays is limited to reactions that occur in aqueous solutions. Here we report an arrayed lipid bilayer chamber system (ALBiC) that contains sub-million femtolitre chambers, each sealed with a stable 4-μm-diameter lipid bilayer membrane. When reconstituted with a limiting amount of the membrane transporter proteins α-hemolysin or F0F1-ATP synthase, the chambers within the ALBiC exhibit stochastic and quantized transporting activities. This demonstrates that the single-molecule analysis of passive and active membrane transport is achievable with the ALBiC system. This new platform broadens the versatility of femtolitre chamber arrays and paves the way for novel applications aimed at furthering our mechanistic understanding of membrane proteins’ function.

Suggested Citation

  • Rikiya Watanabe & Naoki Soga & Daishi Fujita & Kazuhito V. Tabata & Lisa Yamauchi & Soo Hyeon Kim & Daisuke Asanuma & Mako Kamiya & Yasuteru Urano & Hiroaki Suga & Hiroyuki Noji, 2014. "Arrayed lipid bilayer chambers allow single-molecule analysis of membrane transporter activity," Nature Communications, Nature, vol. 5(1), pages 1-8, December.
    • Handle: RePEc:nat:natcom:v:5:y:2014:i:1:d:10.1038_ncomms5519
    DOI: 10.1038/ncomms5519
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