Engineered Proteins for Nanotechnology Applications

描述（由申请人提供）：本提案的目的是设计和生产用于纳米技术、生物材料和传感器应用的工程蛋白质。我们的目标是开发一套分子组件，允许系统组装的功能性纳米结构的分子“支柱”和“节点”。支柱是基本上用作线性结构元件的分子组件。节点是具有多个附着点的分子连接器，这些附着点具有已定义的几何体。支柱和节点的组合导致产生2-D和3-D晶格，其本身可以具有效用和/或可以通过化学修饰或掺入另外的特异性结合蛋白进一步官能化。我们最初的支柱分子是链霉亲和素，一种具有D2对称性的MW 60，000的四聚体蛋白质分子，具有4个维生素生物素的高亲和力结合位点。在支柱应用中，每个链霉亲和素四聚体将与节点蛋白形成2个生物素结合相互作用。当前提案的初始目标是设计和产生两个节点蛋白：一个具有三重对称性以允许形成自组织2D六边形晶格，另一个具有四重对称性以允许形成自组织2D正方形晶格。我们已经从来自耐热细菌的蛋白质的已知晶体结构中选择了候选节点分子，并提出使用蛋白质工程方法来a）。引入可以用生物素官能化的位点，用于与支柱蛋白链霉亲和素B连接。）引入锚定序列（例如聚组氨酸序列）和/或可以用其他化学分子（例如脂质或硫醇）官能化的位点，以允许在2D表面上定向固定，以及c）产生具有降低的结合位点对称性的单链变体，其可以用于顺序组装结构复杂的纳米结构。我们打算在Imiplex公司设计分子组件和编码基因，通过合同生物技术公司生产它们，并通过我们自己的资源和与大学研究实验室的合作关系对其进行评估。我们打算通过直接的基于网络的销售过程，使这些材料广泛提供给生物医学，生物技术和纳米纤维社区。这些材料的可用性将促进新一代功能纳米器件、生物传感器和生物材料的开发。

DESCRIPTION (provided by applicant): The aim of this proposal is the design and production of engineered proteins for nanotechnology, biomaterials and sensor applications. Our objective is to develop a set of molecular components allowing the systematic assembly of functional nanostructures constructed of molecular "struts" and "nodes". Struts are molecular components that basically function as linear structural elements. Nodes are molecular connectors that have multiple attachment points with defined geometry. The combination of struts and nodes results in the production of 2 and 3-D lattices that can have utility themselves and/or can be further functionalized through chemical modification or the incorporation of additional specific binding proteins. Our initial strut molecule is streptavidin, a tetrameric protein molecule of MW 60,000 with D2 symmetry that has 4 high-affinity binding sites for the vitamin biotin. In the strut application, each streptavidin tetramer will form 2 biotin-binding interactions with a node protein. The initial objective of the current proposal is to design and produce two node proteins: one having three-fold symmetry to allow the formation of self-organizing 2D hexagonal lattices, and a node with 4-fold symmetry allowing formation of self-organizing 2D square lattices. We have selected candidate node molecules from known crystal structures of proteins derived from thermostable bacteria, and propose to use protein engineering approaches to a.) introduce sites that can be functionalized with biotin for connection with the strut protein streptavidin b.) introduce anchoring sequences (e.g. poly-histidine sequences) and/or sites that can be functionalized with other chemical molecules (e.g. lipids or thiols) to allow oriented immobilization on 2D surfaces, and c) create single chain variants with reduced binding site symmetry that can be used to sequentially assemble structurally complex nanostructures. We intend to design the molecular components and the encoding genes at our company Imiplex, to produce them through a contract biotechnology company, and to evaluate them through our own resources and collaborative relationships with university research laboratories. We intend to make these materials widely available to the biomedical, biotechnology and nanofabrication communities through a direct web-based sales process. Availability of these materials will facilitate the development of a new generation of functional nanodevices, biosensors, and biomaterials.