Engineered Proteins for Nanotechnology Applications

描述(申请人提供)：该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，允许系统地组装由分子“支柱”和“节点”构成的功能纳米结构。Struts是分子组件，其基本功能是线性结构元素。节点是具有多个具有定义几何图形的附着点的分子连接器。Struts和Node的组合产生了2-D和3-D晶格，这些晶格本身具有用途和/或可以通过化学修饰或加入额外的特定结合蛋白来进一步功能化。我们最初的支柱分子是链霉亲和素，这是一种分子量为60,000的四聚体蛋白质分子，具有D2对称性，与维生素生物素有4个高亲和力结合位点。在STRUT应用中，每个链霉亲和素四聚体将与一个节点蛋白形成两个生物素结合的相互作用。目前的方案的最初目标是设计和生产两个节点蛋白质：一个具有三重对称性，允许形成自组织的二维六角形晶格，以及一个具有四重对称性的节点，允许形成自组织的二维方晶格。我们已经从已知的耐热细菌蛋白质的晶体结构中选择了候选节点分子，并建议使用蛋白质工程方法来实现。)介绍可以用生物素功能化的位点，以便与支柱蛋白链霉亲和素b连接。)引入锚定序列(例如多组氨酸序列)和/或可以与其他化学分子(例如脂类或硫醇)功能化的位点，以允许在2D表面上定向固定，以及c)产生结合位点对称性降低的单链变体，其可用于顺序组装结构复杂的纳米结构。我们打算在我们的公司Imiplex设计分子成分和编码基因，通过一家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它们进行评估。我们打算通过基于网络的直接销售过程，向生物医学、生物技术和纳米制造社区广泛提供这些材料。这些材料的可获得性将促进新一代功能纳米器件、生物传感器和生物材料的发展。

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.