Sweeney Rozamond Y, Park Eun Young, Iverson Brent L, Georgiou George
Institute for Cellular and Molecular Biology, University of Texas, 1 University Station, Austin, 78712, USA.
Biotechnol Bioeng. 2006 Oct 20;95(3):539-45. doi: 10.1002/bit.20886.
One barrier to the construction of nanoscale devices is the ability to place materials into 2D- and 3D-ordered arrays by controlling the assembly and ordering of connections between nanomaterials. Ordered assembly of nanoscale materials may potentially be achieved using biological tools that direct specific connections between individual components. Recently, viruses were successfully employed as scaffolds for the nucleation of nanoparticles and nanowires (Mao et al., 2004); however, there is a paucity of methods for the higher order assembly of phage-templated materials. Here we describe a general strategy for the assembly of filamentous bacteriophages into long, wire-like or into tripod-like structures. To prepare the linear phage assemblies, dimeric leucine zipper protein domains, fused to the p3 and p9 proteins of M13 bacteriophage, were employed to direct the specific end-to-end self-association of the bacteriophage particles. Electron microscopy revealed that up to 90% of the phage displaying complementary leucine zipper domains formed linear multi-phage assemblies, composed of up to 30 phage in length. To prepare tripod-like assemblies, phage were engineered to express trimeric leucine zippers as p3 fusion proteins. This resulted in 3D assembly with three individual phages attached at a single point. These ordered phage structures should provide a foundation for self-assembly of virally templated nanomaterials into useful devices.
构建纳米级器件的一个障碍是能否通过控制纳米材料之间连接的组装和排序，将材料排列成二维和三维有序阵列。利用能够指导单个组件之间特定连接的生物工具，有可能实现纳米级材料的有序组装。最近，病毒已成功用作纳米颗粒和纳米线成核的支架（Mao等人，2004年）；然而，用于噬菌体模板材料高阶组装的方法却很少。在此，我们描述了一种将丝状噬菌体组装成长丝状或三脚架状结构的通用策略。为了制备线性噬菌体组装体，将与M13噬菌体的p3和p9蛋白融合的二聚亮氨酸拉链蛋白结构域用于指导噬菌体颗粒的特定端对端自组装。电子显微镜显示，高达90%展示互补亮氨酸拉链结构域的噬菌体形成了线性多噬菌体组装体，其长度由多达30个噬菌体组成。为了制备三脚架状组装体，对噬菌体进行改造，使其表达作为p3融合蛋白的三聚亮氨酸拉链。这导致了三维组装，三个单独的噬菌体在一个点上相连。这些有序的噬菌体结构应为将病毒模板化的纳米材料自组装成有用的器件奠定基础。
