ORIENTED PROTEIN ARRAYS ON POLYMERS NANOSTRUCTURES

聚合物纳米结构上的定向蛋白质阵列

• 批准号: 6385200
• 负责人: Angelika Niemz
• 金额: $ 2.49万
• 依托单位: CALIFORNIA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-06-01 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6385200
• 关键词: active sites; biomaterial development /preparation; biosensor device; crosslink; cytochrome c; electrochemistry; electron transport; macromolecule; microcapsule; nanotechnology; peptide chemical synthesis; polymerization; polymers

A novel technique is presented for engineering two dimensional, oriented, and functional protein arrays immobilized on polymer nanospheres, templated through appropriately functionalized liposomes. Cytochrome c (cyt c) adsorbs on anionic lipid bilayers via the patch of charged residues responsible for precursory complex formation during electron transfer. Cyt c arrays bound to the inside wall of large unilamellar vesicles will be immobilized through photo initiated polymerization of an encapsulated prepolymer and covalent crosslinking of the protein to the polymer matrix. Removal of the lipid bilayer will release the nanosphere and expose the functional protein surface area. The biological activity of these cyt c nanostructures will be established through a variety of methods, which will also explore their applicability as mediators in electron transfer biosensors, and as affinity probes for electron transfer proteins, and their potential to provide new insight into the macromolecular recognition processes essential for regulating biological electron transfer. The immobilization techniques presented herein can be applied to other proteins which self- assemble on lipid bilayers with specific orientation due to ligand-binding or electrostatic interactions. It is a simple, general method to obtain functional, oriented protein arrays.

提出了一种新的技术，用于工程二维，定向和功能蛋白阵列固定在聚合物纳米球上，并通过适当功能化的脂质体模板。细胞色素c（Cyt C）吸附在阴离子脂质双层上，通过负责在电子传递期间负责前保体复合物形成的带电残基。通过照片启动的聚合聚合的聚合和蛋白质与聚合物基质的共价交联，将固定在大型单层囊泡内壁上的Cyt C阵列。去除脂质双层将释放纳米球并暴露功能性蛋白质表面积。这些Cyt C纳米结构的生物学活性将通过多种方法建立，这还将探索它们作为电子传输生物传感器中介体的适用性，并作为电子传递蛋白的亲和力探针，以及它们为调节生物学电子传递所必需的新洞察力提供新的洞察力。本文介绍的固定技术可以应用于其他蛋白质，这些蛋白质会因配体结合或静电相互作用而在脂质双层中自我组装。这是一种获得功能，定向蛋白质阵列的简单，通用的方法。