Department of Material Science and Engineering, Genetically Engineered Materials Science and Engineering Center (GEMSEC), University of Washington, Seattle, Washington 98195, USA.
Biotechnol Bioeng. 2012 May;109(5):1120-30. doi: 10.1002/bit.24405. Epub 2011 Dec 26.
Development of versatile and flexible assembly systems for fabrication of functional hybrid nanomaterials with well-defined hierarchical and spatial organization is of a significant importance in practical nanobiotechnology applications. Here we demonstrate a bio-enabled self-assembly technique for fabrication of multi-layered protein and nanometallic assemblies utilizing a modular gold-binding (AuBP1) fusion tag. To accomplish the bottom-up assembly we first genetically fused the AuBP1 peptide sequence to the C'-terminus of maltose-binding protein (MBP) using two different linkers to produce MBP-AuBP1 hetero-functional constructs. Using various spectroscopic techniques, surface plasmon resonance (SPR) and localized surface plasmon resonance (LSPR), we verified the exceptional binding and self-assembly characteristics of AuBP1 peptide. The AuBP1 peptide tag can direct the organization of recombinant MBP protein on various gold surfaces through an efficient control of the organic-inorganic interface at the molecular level. Furthermore using a combination of soft-lithography, self-assembly techniques and advanced AuBP1 peptide tag technology, we produced spatially and hierarchically controlled protein multi-layered assemblies on gold nanoparticle arrays with high molecular packing density and pattering efficiency in simple, reproducible steps. This model system offers layer-by-layer assembly capability based on specific AuBP1 peptide tag and constitutes novel biological routes for biofabrication of various protein arrays, plasmon-active nanometallic assemblies and devices with controlled organization, packing density and architecture.
开发多功能、灵活的组装系统，用于制造具有明确的层次和空间组织的功能混合纳米材料，在实际的纳米生物技术应用中具有重要意义。在这里，我们展示了一种利用模块化金结合（AuBP1）融合标签制造多层蛋白质和纳米金属组装体的生物启发自组装技术。为了实现自下而上的组装，我们首先使用两种不同的接头将 AuBP1 肽序列基因融合到麦芽糖结合蛋白（MBP）的 C'-末端，生成 MBP-AuBP1 杂功能构建体。我们使用各种光谱技术，如表面等离子体共振（SPR）和局域表面等离子体共振（LSPR），验证了 AuBP1 肽的特殊结合和自组装特性。AuBP1 肽标签可以通过在分子水平上高效控制有机-无机界面，指导重组 MBP 蛋白在各种金表面上的组织。此外，我们还结合软光刻、自组装技术和先进的 AuBP1 肽标签技术，在金纳米粒子阵列上产生了具有高分子堆积密度和图案化效率的空间和层次控制的蛋白质多层组装体，这些组装体具有简单、可重复的步骤。该模型系统提供了基于特定 AuBP1 肽标签的逐层组装能力，并为各种蛋白质阵列、等离子体活性纳米金属组装体和具有受控组织、堆积密度和结构的设备的生物制造构成了新的生物学途径。
