title: Arrayed lipid bilayer chambers allow single-molecule analysis of membrane transporter activity.
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.
authors: 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].
authors: Rikiya
authors: R
authors: Watanabe
authors: 1] Department of Applied Chemistry, Graduate School of Engineering, The University of Tokyo, Bunkyo-ku, Tokyo 113-8656, Japan [2].
authors: Naoki
authors: N
authors: Soga
authors: 1] Department of Applied Chemistry, Graduate School of Engineering, The University of Tokyo, Bunkyo-ku, Tokyo 113-8656, Japan [2].
authors: Daishi
authors: D
authors: Fujita
authors: 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.
authors: Kazuhito V
authors: KV
authors: Tabata
authors: 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.
authors: Lisa
authors: L
authors: Yamauchi
authors: 1] CREST, JST, Bunkyo-ku, Tokyo 113-8656, Japan [2] Institute of Industrial Science, The University of Tokyo, Meguro-ku, Tokyo 153-8505, Japan.
authors: Soo
authors: S
authors: Hyeon Kim
authors: Laboratory of Neurobiology, Graduate School of Medicine, The University of Tokyo, Bunkyo-ku, Tokyo 113-0033, Japan.
authors: Daisuke
authors: D
authors: Asanuma
authors: Laboratory of Chemical Biology and Molecular Imaging, Graduate School of Medicine, The University of Tokyo, Bunkyo-ku, Tokyo 113-0033, Japan.
authors: Mako
authors: M
authors: Kamiya
authors: Laboratory of Chemical Biology and Molecular Imaging, Graduate School of Medicine, The University of Tokyo, Bunkyo-ku, Tokyo 113-0033, Japan.
authors: Yasuteru
authors: Y
authors: Urano
authors: 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.
authors: Hiroaki
authors: H
authors: Suga
authors: 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.
authors: Hiroyuki
authors: H
authors: Noji
mesh_terms: D001427:Bacterial Toxins / Q000378:metabolism; D001693:Biological Transport, Active; D005456:Fluorescent Dyes / Q000378:metabolism; D006460:Hemolysin Proteins / Q000378:metabolism; D006834:Hydrazines / Q000378:metabolism; D008051:Lipid Bilayers / Q000378:metabolism*; D026901:Membrane Transport Proteins / Q000032:analysis / Q000378:metabolism*; D055616:Microtechnology; D008967:Molecular Biology / Q000295:instrumentation* / Q000379:methods*; D036103:Nanotechnology / Q000295:instrumentation / Q000379:methods; D038761:Photobleaching; D006180:Proton-Translocating ATPases / Q000378:metabolism
chemical_list: C417561:Alexa 488 hydrazide; D001427:Bacterial Toxins; D005456:Fluorescent Dyes; D006460:Hemolysin Proteins; D006834:Hydrazines; D008051:Lipid Bilayers; D026901:Membrane Transport Proteins; C021091:staphylococcal alpha-toxin; D006180:Proton-Translocating ATPases