Format

Send to

Choose Destination
Nat Commun. 2014 Jul 24;5:4519. doi: 10.1038/ncomms5519.

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

Author information

1
1] Department of Applied Chemistry, Graduate School of Engineering, The University of Tokyo, Bunkyo-ku, Tokyo 113-8656, Japan [2] PRESTO, JST, Bunkyo-ku, Tokyo 113-8656, Japan [3] CREST, JST, Bunkyo-ku, Tokyo 113-8656, Japan [4].
2
1] Department of Applied Chemistry, Graduate School of Engineering, The University of Tokyo, Bunkyo-ku, Tokyo 113-8656, Japan [2].
3
1] Department of Applied Chemistry, Graduate School of Engineering, The University of Tokyo, Bunkyo-ku, Tokyo 113-8656, Japan [2] PRESTO, JST, Bunkyo-ku, Tokyo 113-8656, Japan [3] CREST, JST, Bunkyo-ku, Tokyo 113-8656, Japan.
4
1] Department of Applied Chemistry, Graduate School of Engineering, The University of Tokyo, Bunkyo-ku, Tokyo 113-8656, Japan [2] CREST, JST, Bunkyo-ku, Tokyo 113-8656, Japan.
5
1] CREST, JST, Bunkyo-ku, Tokyo 113-8656, Japan [2] Institute of Industrial Science, The University of Tokyo, Meguro-ku, Tokyo 153-8505, Japan.
6
Laboratory of Neurobiology, Graduate School of Medicine, The University of Tokyo, Bunkyo-ku, Tokyo 113-0033, Japan.
7
Laboratory of Chemical Biology and Molecular Imaging, Graduate School of Medicine, The University of Tokyo, Bunkyo-ku, Tokyo 113-0033, Japan.
8
1] CREST, JST, Bunkyo-ku, Tokyo 113-8656, Japan [2] Department of Chemistry, Graduate School of Science, The University of Tokyo, Bunkyo-ku, Tokyo 113-8656, Japan.

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.

PMID:
25058452
PMCID:
PMC4124872
DOI:
10.1038/ncomms5519
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for Nature Publishing Group Icon for PubMed Central
Loading ...
Support Center