Engineered Macromolecule for Controlled Synthesis in Nanotechnology

描述（由申请人提供）：我们的目标是开发一种链霉亲和素大分子衔接子（SAMA），能够控制链霉亲和素连接的蛋白质缀合物的组装，用于纳米技术、生物材料和传感器应用。链霉亲和素（SAV）是具有D2对称性的MW 60，000的四聚体蛋白质分子，其具有4个针对维生素生物素的高亲和力结合位点。由于链霉亲和素-生物素复合物的高稳定性，再加上通过定向化学修饰使蛋白质生物素化的能力，链霉亲和素在生物技术应用中得到了广泛的应用，并且也是基于蛋白质的纳米技术中的关键构件。在使用链霉亲和素-生物素相互作用的许多应用中，由于缺乏对大分子缀合和组装过程的控制，所得产物的质量和均一性受到限制。本提案描述了基于热稳定二聚体蛋白质MJ 0557的工程化形式的链霉亲和素大分子衔接子（SAMA）的开发。设想SAMA有效地用作固定剂和可逆保护基团，提供从固相化学合成已知的关键优点：1）反应性的几何控制，2）生长分子组装体的特异性固定的潜力，3）使用质量作用驱动反应平衡完成的能力，和4）大大促进从试剂中纯化反应产物的能力。该项目的具体目标涉及SAMA和SAV：SAMA复合物的详细设计、表达和生物物理表征。我们打算通过基于网络的销售和分销，将SAV：SAMA复合物广泛提供给生物医学，生物技术和纳米纤维社区。我们相信，这种SAMA的普遍可用性将促进新一代功能性纳米器件、生物传感器和生物材料的开发，从而构成疾病诊断和皮肤、骨骼和其他组织替代材料的新方法。我们相信，指导含链霉亲和素的组装体的受控合成的分子的普遍可用性将极大地促进新一代功能性纳米器件、生物传感器和生物材料的开发，这些功能性纳米器件、生物传感器和生物材料进而构成用于疾病诊断和皮肤、骨骼和其他组织的替代材料的新方法。

DESCRIPTION (provided by applicant): Our objective is to develop a Streptavidin Macromolecular Adaptor (SAMA) enabling the controlled assembly of streptavidin-linked protein conjugates for nanotechnology, biomaterials and sensor applications. Streptavidin (SAV) is a tetrameric protein molecule of MW 60,000 with D2 symmetry that has 4 high-affinity binding sites for the vitamin biotin. Owing to the high stability of the streptavidin-biotin complex, coupled with the ability to biotinylate proteins through directed chemical modification, streptavidin has found wide use in biotechnology applications and has also been a key building block in protein-based nanotechnology. In many applications where streptavidin-biotin interactions are used, the quality and homogeneity of the resulting product is limited owing to lack of control over the macromolecular conjugation and assembly process. The present proposal describes the development of a Streptavidin Macromolecular Adaptor (SAMA), based on an engineered form of a thermostable dimeric protein MJ0557. SAMA is envisioned to effectively function both as an immobilization agent and as a reversible protecting group providing key advantages known from solid-phase chemical synthesis: 1) geometrical control of reactivity, 2) potential for specific immobilization of a growing molecular assembly, 3) ability to drive reaction equilibria to completion using mass action, and 4) greatly facilitated ability to purify reaction products from reagents. The specific aims of the project involve the detailed design, expression and biophysical characterization of SAMA and the SAV:SAMA complex. We intend to make the SAV:SAMA complex widely available to the biomedical, biotechnology and nanofabrication communities through web-based sales and distribution. We believe that the general availability of this SAMA will facilitate the development of a new generation of functional nanodevices, biosensors, and biomaterials, that in turn constitute new approaches for disease diagnostics and replacement materials for skin, bones and other tissues. We believe that the general availability of a molecule to direct the controlled synthesis of streptavidin- containing assemblies will tremendously facilitate the development of a new generation of functional nanodevices, biosensors, and biomaterials that in turn constitute new approaches for disease diagnostics and replacement materials for skin, bones and other tissues.