Generic Assays for Functional Characterization (548-591)

功能表征的通用分析 (548-591)

• 批准号: 9050048
• 负责人: WAYNE A. HENDRICKSON
• 金额: $ 40.73万
• 依托单位: NEW YORK STRUCTURAL BIOLOGY CENTER
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-05-01 至 2021-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9050048
• 关键词: Affinity; Binding; Biological Assay; Cell physiology; Detergents; Development; Devices; Environment; Enzymes; Evaluation; Event; Generic Drugs; High Pressure Liquid Chromatography; Homeostasis; Hydrogels; Ion Channel; Kinetics; Label; Life; Ligands; Lipid Bilayers; Lipids; Liposomes; Maintenance; Measures; Membrane; Membrane Proteins; Membrane Transport Proteins; Metals; Methods; Micelles; Monitor; Noise; Play; Preparation; Process; Production; Proteins; Protocols documentation; Radioactive; Research; Resources; Roentgen Rays; Role; Sampling; Semiconductors; Signal Transduction; System; Techniques; Technology; Testing; Vesicle; Work; experience; high throughput screening; improved; instrument; light scattering; novel; nutrition; protein reconstitution; proteoliposomes; screening; temporal measurement; uptake

Membrane proteins play critical roles in virtually all life processes. Membrane proteins regulate cellular processes by functioning as gates for signal transduction, nutrition transport, defense from outside attack, and maintenance of membrane homeostasis. However, due to difficulties in handling membrane proteins and the requirement of a lipid-bilayer environment, functional studies are never straightforward and require substantial efforts in labor-intensive screening for conditions suitable for functional characterization. To accelerate functional studies of membrane proteins, we propose to develop a technical research resource for generic functional characterization of membrane proteins. Built on our nearly ten years of research experiences on membrane proteins and our functional characterization of more than a dozen novel membrane transporters, channels and enzymes, we propose to develop complementary and synergic techniques in three sub-projects. Aim 1 (TR&D3.1) focuses on the development of microscale thermophoresis technology for analysis of the interactions, dynamics and kinetics of membrane proteins. Aim 2 (TR&D3.2) combines the complementary metaloxide- semiconductor (CMOS) chips with the planar lipid bilayer membranes for recording membrane channels. Aim 3 (TR&D3.3) utilizes fluorescent and radioactive labels for probing uptake activities for membrane transporters and channels. The three integrated technologies will be robust and transformative for characterization of challenging membrane proteins.

膜蛋白几乎在所有生命过程中都发挥着关键作用。膜蛋白调节细胞 通过充当信号转导、营养运输、防御外界攻击的大门，以及 维持膜稳态。然而，由于处理膜蛋白的困难和 由于需要脂质双层环境，功能研究从来都不是简单的，需要大量的 努力进行劳动密集型筛选，寻找适合功能表征的条件。加速 膜蛋白的功能研究，我们建议开发通用技术研究资源 膜蛋白的功能表征。基于我们近十年的研究经验 膜蛋白和我们对十多种新型膜转运蛋白的功能表征， 通道和酶，我们建议在三个子项目中开发互补和协同技术。 目标 1 (TR&D3.1) 重点开发用于分析的微尺度热泳技术 膜蛋白的相互作用、动力学和动力学。目标 2 (TR&D3.2) 结合了互补的金属氧化物- 具有用于记录膜通道的平面脂质双层膜的半导体（CMOS）芯片。目标 3 (TR&D3.3) 利用荧光和放射性标记来探测膜转运蛋白的摄取活性和 渠道。这三种集成技术对于表征具有挑战性的技术将具有强大的变革性 膜蛋白。