S-layer-streptavidin fusion proteins as template for nanopatterned molecular arrays
S-layer-streptavidin fusion proteins as template for nanopatterned molecular arrays
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DOI：
10.1073/pnas.232299399
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发表时间：
2002-11-12
影响因子：
11.1
通讯作者：
Sára, M
中科院分区：
文献类型：
--
作者：
Moll, D;Huber, C;Sára, M
生物分子自组装可以作为一个强有力的工具，纳米工程。在本文中，我们描述了纳米生物技术，这是基于链霉亲和素的融合到一个结晶的细菌细胞表面层（S-层）蛋白质的固有能力，自组装成一个单分子的蛋白质晶格的积木的发展。在大肠杆菌中独立地产生融合蛋白和链霉亲和素，分离并混合以重折叠和纯化1：3化学计量的异源四聚体。自组装嵌合S层可以在悬浮液中、脂质体上、硅片上和含有细胞壁片段的辅助细胞壁聚合物上形成。二维蛋白质晶体显示链霉亲和素在定义的重复间隔，并且它们能够结合D-生物素和生物素化的蛋白质。因此，嵌合S层可以用作自组装纳米图案化分子亲和基质以在表面上排列生物素化化合物。此外，它还具有作为脂质体功能性包衣的应用潜力。
Biomolecular self-assembly can be used as a powerful tool for nanoscale engineering. In this paper, we describe the development of building blocks for nanobiotechnology, which are based on the fusion of streptavidin to a crystalline bacterial cell surface layer (S-layer) protein with the inherent ability to self-assemble into a monomolecular protein lattice. The fusion proteins and streptavidin were produced independently in Escherichia coli, isolated, and mixed to refold and purify heterotetramers of 1:3 stoichiometry. Self-assembled chimeric S-layers could be formed in suspension, on liposomes, on silicon wafers, and on accessory cell wall polymer containing cell wall fragments. The two-dimensional protein crystals displayed streptavidin in defined repetitive spacing, and they were capable of binding D-biotin and biotinylated proteins. Therefore, the chimeric S-layer can be used as a self-assembling nanopatterned molecular affinity matrix to arrange biotinylated compounds on a surface. In addition, it has application potential as a functional coat of liposomes.