Hybrid Model Membrane Combining Micropatterned Lipid Bilayer and Hydrophilic Polymer Brush
Hybrid Model Membrane Combining Micropatterned Lipid Bilayer and Hydrophilic Polymer Brush
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结合微图案脂质双层和亲水聚合物刷的混合模型膜
DOI：
10.1021/acs.langmuir.7b00463
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发表时间：
2017
期刊：
影响因子：
3.9
通讯作者：
Morigaki Kenichi
中科院分区：
文献类型：
--
作者：
Nishimura Toshiki;Tamura Fuyuko;Kobayashi Sawako;Tanimoto Yasushi;Hayashi Fumio;Sudo Yuki;Iwasaki Yasuhiko;Morigaki Kenichi
基底支持的平面脂质双层（SPB）被用作生物膜的多功能模型系统。然而，固体支持物和膜之间的接近限制了 SPB 在膜蛋白功能分析中的实用性。在这里，我们提出了一种模型膜，通过将聚合脂质双层的稳定支架与亲水性聚合物刷相结合，可以扩大基底表面和膜之间的距离。通过丁二炔脂质的光刻聚合以及随后掺入天然（流体）脂质双层来生成微图案 SPB。在嵌入液体脂质双层的情况下，通过水溶液中的原位原子转移自由基聚合（ATRP）在聚合物双层表面形成聚-2-甲基丙烯酰氧基乙基磷酰胆碱（poly(MPC)）的亲水性聚合物刷。模型膜蛋白（Haloquadratum walsbyibacteriorhodopsin：HwBR）可以重构为聚合物刷支持的双层，其中固定分子显着减少。此外，聚合物刷端子可以通过连续聚合MPC和甲基丙烯酸2-氨基乙酯（AMA）来功能化。聚 (AMA) 的反应性胺部分能够将多种生物分子和表面缀合到膜上。微图案双层和聚合物刷的组合模仿了生物膜的二维和三维结构，为在真正的仿生环境中测定膜蛋白提供了一个平台。
Substrate-supported planar lipid bilayers (SPBs) are being utilized as a versatile model system of the biological membrane. However, the proximity between the solid support and membrane limits utility of SPBs for the functional analyses of membrane proteins. Here, we present a model membrane that can enlarge the distance between the substrate surface and the membrane by combining a stable scaffold of polymerized lipid bilayer with a hydrophilic polymer brush. A micropatterned SPB was generated by the lithographic polymerization of diacetylene lipids and subsequent incorporation of natural (fluid) lipid bilayers. Hydrophilic polymer brush of poly-2-methacryloyloxyethyl phosphorylcholine (poly(MPC)) was formed on the surface of polymeric bilayer by thein situatom transfer radical polymerization (ATRP) in aqueous solution, in the presence of embedded fluid lipid bilayers. A model membrane protein (Haloquadratum walsbyibacteriorhodopsin: HwBR) could be reconstituted into the polymer brush-supported bilayers with significantly reduced immobile molecules. Furthermore, the polymer brush terminals could be functionalized by successively polymerizing MPC and 2-aminoethyl methacrylate (AMA). The reactive amine moiety of poly(AMA) enables to conjugate a wide range of biological molecules and surfaces to the membrane. The combination of micropatterned bilayer and polymer brush mimics the two- and three-dimensional structures of the biological membrane, providing a platform to assay membrane proteins in a truly biomimetic environment.