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首页> 外文期刊>Advanced Functional Materials >Supported Cell-Membrane Sheets for Functional Fluorescence Imaging of Membrane Proteins
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Supported Cell-Membrane Sheets for Functional Fluorescence Imaging of Membrane Proteins

机译：支持的膜蛋白功能性荧光成像的细胞膜片

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摘要

Cell-membrane sheets suitable for in-vitro functional fluorescence studies have been prepared by direct detachment from cell membranes using poly-L-lysine-coated glass slides. The resulting transferred planar membranes conserve the composition as well as most properties of the original plasma membrane; in particular, both membrane leaflets remain fluid, allowing the investigation of diffusion properties of different cellular membrane components. Measurements on membrane sheets offer several advantages as compared to those on living cells. First, access to the intracellular leaflet is obtained, in particular to the intra-cellular part of membrane proteins and to cytoplasmic membrane-associated proteins, opening the possibility of labeling them and modulating their properties with membrane impermeable compounds. Second, the cytosolic autofluorescence of the cells is absent, allowing ultrasensitive measurements to be performed down to the single-molecule level. Third, the complexity of cellular processes occurring at the plasma membrane can be reduced, allowing the sequential investigation of selected events from complex biochemical networks. These advantages are illustrated by ligand-binding studies on the α1b-adrenergic receptor. Our results indicate that supported membrane sheets might find a broad application as an ideal in-vitro system for the elucidation of complex signaling pathways.
机译：通过使用聚-L-赖氨酸涂层的载玻片直接从细胞膜上分离，已经制备了适合体外功能荧光研究的细胞膜片。所得转移的平面膜可保留原始质膜的组成以及大多数特性；特别地，两个膜小叶都保持流体状态，从而允许研究不同细胞膜组分的扩散特性。与在活细胞上进行的测量相比，在膜片上进行的测量具有许多优势。首先，获得进入细胞内小叶的途径，特别是进入膜蛋白的细胞内部分以及与细胞质膜相关的蛋白，这为标记它们和用膜不透性化合物调节其性质提供了可能性。其次，不存在细胞的胞质自发荧光，从而可以进行超灵敏的测量，直至单分子水平。第三，可以降低在质膜上发生的细胞过程的复杂性，从而允许对来自复杂生化网络的选定事件进行顺序研究。这些优势通过对α1b-肾上腺素受体的配体结合研究得以说明。我们的结果表明，支持的膜片可以作为阐明复杂信号通路的理想的体外系统找到广泛的应用。

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