Engineered Macromolecule for Controlled Synthesis in Nanotechnology

描述(申请人提供)：我们的目标是开发一种链霉亲和素大分子适配器(SAMA)，使链霉亲和素连接的蛋白质偶联物能够在纳米技术、生物材料和传感器应用中进行受控组装。链霉亲和素(SAV)是一种分子量为60,000、D2对称性的四聚体蛋白质分子，与维生素生物素有4个高亲和力结合位点。由于链霉亲和素-生物素复合体的高度稳定性，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中得到了广泛的应用，也成为基于蛋白质的纳米技术的关键构件。在许多使用链霉亲和素-生物素相互作用的应用中，由于缺乏对大分子连接和组装过程的控制，最终产品的质量和均质性受到限制。本提案描述了基于耐热二聚体蛋白MJ0557的工程形式的链霉亲和素大分子接头(SAMA)的开发。SAMA有望有效地作为固定剂和可逆保护基团发挥作用，提供固相化学合成的关键优点：1)反应活性的几何控制，2)特定固定不断增长的分子组装的潜力，3)通过质量作用驱动反应平衡完成的能力，以及4)极大地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复合体的详细设计、表达和生物物理特性。我们打算通过基于网络的销售和分销，向生物医学、生物技术和纳米制造社区广泛提供SAV：SAMA复合体。我们相信，这种SAMA的全面上市将促进新一代功能纳米设备、生物传感器和生物材料的开发，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。我们相信，指导含有链霉亲和素的组件的受控合成的分子的普遍可获得性将极大地促进新一代功能纳米设备、生物传感器和生物材料的开发，这些设备和材料反过来又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.