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.

膜蛋白在几乎所有的生命过程中起着关键作用。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， 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 在劳动密集型筛选适合功能表征的条件方面的努力。加快 膜蛋白的功能研究，我们建议开发一个通用的技术研究资源， 膜蛋白的功能表征。基于我们近十年的研究经验， 膜蛋白和我们对十几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）侧重于开发用于分析生物样品的微尺度热泳技术。 膜蛋白的相互作用、动力学和动力学。目标2（TR&D3.2）结合互补金属氧化物- 图1示出了具有用于记录膜通道的平面脂质双层膜的CMOS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， 渠道这三种集成技术将是强大的和变革性的表征具有挑战性的 膜蛋白