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.

提出了一种新技术，用于设计固定在聚合物纳米球上的二维、定向和功能性蛋白质阵列，通过适当功能化的脂质体作为模板。细胞色素 c (cyt c) 通过负责电子转移过程中前体复合物形成的带电残基斑块吸附在阴离子脂质双层上。与大单层囊泡内壁结合的细胞色素 c 阵列将通过封装预聚物的光引发聚合以及蛋白质与聚合物基质的共价交联来固定。去除脂质双层将释放纳米球并暴露功能性蛋白质表面积。这些细胞色素c纳米结构的生物活性将通过多种方法确定，这还将探索它们作为电子转移生物传感器中的介体和作为电子转移蛋白的亲和探针的适用性，以及它们为大分子识别提供新见解的潜力调节生物电子转移所必需的过程。本文提出的固定技术可应用于由于配体结合或静电相互作用而在脂质双层上以特定方向自组装的其他蛋白质。这是一种获得功能性、定向蛋白质阵列的简单、通用的方法。