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钝端 DNA 瓦片在支撑脂质双层上的长程有序排列。

Long-Range Ordering of Blunt-Ended DNA Tiles on Supported Lipid Bilayers.

机构信息

Department of Chemistry, McGill University , 801 Sherbrooke Street West, Montreal QC H3A 0B8, Canada.

出版信息

J Am Chem Soc. 2017 Aug 30;139(34):12027-12034. doi: 10.1021/jacs.7b06572. Epub 2017 Aug 21.

Abstract

Long-range ordering of DNA crossover tiles with blunt ends on lipid bilayers is investigated using atomic force microscopy. "Blunt-ended" tiles do not have single-stranded complementary ends, and thus instead of assembling via base-pairing, they can interact by π-stacking of their duplex ends. This work demonstrates that the balance of base π-stacking interactions between the ends of DNA duplexes, cholesterol-mediated DNA anchoring, and electrostatic DNA binding to supported lipid bilayers (SLBs) presents an opportunity to build dynamic materials with long-range order on a soft support. The tiles are shown to organize into novel tunable surface packing morphologies on the micrometer scale. This work focuses on three-point star (3PS) tiles that are either unmodified or modified with a cholesterol unit and investigates their interactions on supported lipid bilayers. On fluid bilayers, the cholesterol tiles form extended hexagonal arrays with few defects, while the unmodified tiles do not bind. In contrast, both modified and unmodified tiles bind to gel-phase bilayers and produce arrays of new organized morphologies. With increasing tile concentration, we observe a range of motifs, that progressively favor tile-tile packing over duplex-end π-stacking. These structures can selectively pattern domains of phase-separated lipid bilayers, and the patterning is also observed for four-arm cross-tiles. Dynamic blunt end contacts promote error correction and network reconfiguration to maximize favorable interactions with the substrate and are required for the observed tile organization. These results suggest that small blunt-ended tiles can be used as a platform to organize oligonucleotides, nanoparticles, and proteins into extensive networks at the interface with biologically relevant membrane systems or other soft surface materials for applications in cellular recognition, plasmonics, light harvesting, model systems for membrane protein assemblies, or analytical devices.

摘要

使用原子力显微镜研究了具有钝末端的 DNA 交叉平铺物在脂质双层上的长程有序排列。“钝末端”平铺物没有单链互补末端,因此它们不是通过碱基配对组装,而是通过双链末端的π堆积相互作用。这项工作表明,DNA 双链末端的碱基π堆积相互作用、胆固醇介导的 DNA 锚定以及静电 DNA 结合到支持的脂质双层(SLB)之间的平衡,为在软支撑上构建具有长程有序的动态材料提供了机会。结果表明,平铺物在微米尺度上组织成新颖的可调表面堆积形态。这项工作集中在三点星(3PS)平铺物上,它们要么未修饰,要么修饰有胆固醇单元,并研究了它们在支持的脂质双层上的相互作用。在流体双层上,胆固醇平铺物形成具有少量缺陷的扩展六边形阵列,而未修饰的平铺物则不结合。相比之下,修饰和未修饰的平铺物都与凝胶相双层结合,并产生新的有组织形态的阵列。随着平铺物浓度的增加,我们观察到一系列图案,这些图案逐渐有利于平铺物之间的堆积而不是双链末端π堆积。这些结构可以选择性地对相分离脂质双层的域进行图案化,并且还观察到四臂交叉平铺物的图案化。动态钝端接触促进错误修正和网络重新配置,以最大化与基底的有利相互作用,这是观察到的平铺物组织所必需的。这些结果表明,小的钝末端平铺物可用作平台,将寡核苷酸、纳米颗粒和蛋白质组织成与生物相关的膜系统或其他软表面材料界面处的广泛网络,用于细胞识别、等离子体学、光捕获、膜蛋白组装的模型系统或分析设备。

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