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


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2014-12

AUTHORS

Rikiya Watanabe, Naoki Soga, Daishi Fujita, Kazuhito V. Tabata, Lisa Yamauchi, Soo Hyeon Kim, Daisuke Asanuma, Mako Kamiya, Yasuteru Urano, Hiroaki Suga, Hiroyuki Noji

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. More... »

PAGES

4519

References to SciGraph publications

Identifiers

URI

http://scigraph.springernature.com/pub.10.1038/ncomms5519

DOI

http://dx.doi.org/10.1038/ncomms5519

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1053025353

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/25058452


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