Urban Michael, Kleefen Alexander, Mukherjee Nobina, Seelheim Patrick, Windschiegl Barbara, Vor der Brüggen Marc, Koçer Armagan, Tampé Robert
Institute of Biochemistry, Biocenter, Goethe-University , Frankfurt, Germany.
Nano Lett. 2014 Mar 12;14(3):1674-80. doi: 10.1021/nl5002873. Epub 2014 Feb 17.
Membrane proteins are prime drug targets as they control the transit of information, ions, and solutes across membranes. Here, we present a membrane-on-nanopore platform to analyze nonelectrogenic channels and transporters that are typically not accessible by electrophysiological methods in a multiplexed manner. The silicon chip contains 250,000 femtoliter cavities, closed by a silicon dioxide top layer with defined nanopores. Lipid vesicles containing membrane proteins of interest are spread onto the nanopore-chip surface. Transport events of ligand-gated channels were recorded at single-molecule resolution by high-parallel fluorescence decoding.
膜蛋白是主要的药物靶点,因为它们控制信息、离子和溶质跨膜转运。在此,我们展示了一种纳米孔上的膜平台,用于以多重方式分析通常无法通过电生理方法检测的非电生性通道和转运体。该硅芯片包含25万个飞升腔,由带有特定纳米孔的二氧化硅顶层封闭。含有感兴趣膜蛋白的脂质囊泡铺展在纳米孔芯片表面。通过高并行荧光解码以单分子分辨率记录配体门控通道的转运事件。