Native Cell Membrane Nanoparticles System

天然细胞膜纳米粒子系统

• 批准号: 10454882
• 负责人: Youzhong Guo
• 金额: $ 38.26万
• 依托单位: VIRGINIA COMMONWEALTH UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-07-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10454882
• 关键词: ATP phosphohydrolase; ATP-Binding Cassette Transporters; Address; Adopted; Bacillus cereus; Bacteria; Biochemical; Biology; Cell Membrane Proteins; Cell membrane; Cells; Characteristics; Cryoelectron Microscopy; Data; Detergents; Environment; Escherichia coli; Goals; Homologous Gene; Human; Insecta; Laboratories; Libraries; Lipid Bilayers; Lipids; Listeria monocytogenes; Maleic Anhydride; Mammalian Cell; Membrane; Membrane Lipids; Membrane Proteins; Membrane Structure and Function; Mind; Modernization; Molecular Weight; NPC1 gene; Niemann-Pick Diseases; Organism; Pharmaceutical Preparations; Play; Polymers; Preparation; Proteins; Protocols documentation; Proton-Translocating ATPases; Research; Resolution; Role; Saccharomyces cerevisiae; Sensory; Structure; Styrenes; System; TRP channel; Testing; Transmembrane Domain; Tryptophan; Work; Yeasts; base; biophysical analysis; cell injury; copolymer; drug development; drug discovery; flexibility; membrane activity; membrane model; nanodisk; nanoparticle; particle; protein structure; protocol development; prototype; screening; structural biology

SUMMARY Membrane proteins play crucial roles in living organisms. Protein-lipid interactions are crucial for the structural and functional integrity of membrane proteins. High-resolution of membrane protein structures are in high-demand for both understanding their biology and structure-based drug discovery. It is often need to extract membrane proteins from cell membranes for biochemical and biophysical study. Currently detergents are predominantly used for membrane protein extraction. Detergents have significant drawbacks in that they destroy lipid bilayer and remove important lipid molecules from membrane proteins. We are developing a detergent-free native cell membrane nanoparticles system for membrane protein research. The native cell membrane nanoparticles system is composed of three components: 1) Membrane active polymers. 2) Protocols for preparation of native cell membrane nanoparticles. 3) Single-particles cryo-EM analysis of the native cell membrane nanoparticles. We propose to develop a system that can be generally applicable to both prokaryotic and eukaryotic membrane proteins. Specific Aim 1 focuses on construction of membrane active polymer library and develop protocols for preparation of native cell membrane nanoparticles using our selected model membrane proteins. We have developed four prototype membrane active polymers, with the similar strategies we plan to expand the membrane active polymers using both commercially available styrene maleic anhydride copolymers and polymers synthesized in laboratories. Specific Ami 2 focuses on developing protocols for high-resolution structure determination of selected prokaryotic model membrane proteins. Specific Aim 3 focuses on developing protocols for high-resolution structures of selected eukaryotic model membrane proteins. The proposed research is significant because it provides a revolutionary detergent-free approach for study membrane proteins in their local lipid environment. As we have demonstrated in our preliminary data, we found the structural information of protein-lipid interaction for membrane proteins is crucial for understanding the structure and function of membrane proteins. This system might transform the membrane protein research field and have a great impact for membrane protein structure based drug discovery and development.

概括 膜蛋白在生物体中起关键作用。蛋白 - 脂质相互作用对于 膜蛋白的结构和功能完整性。膜蛋白结构的高分辨率 高需求既了解它们的生物学和基于结构的药物发现。通常需要 从细胞膜中提取膜蛋白进行生化和生物物理研究。现在 洗涤剂主要用于膜蛋白提取。洗涤剂有明显的缺点 因为它们破坏了脂质双层并从膜蛋白中去除重要的脂质分子。我们是 开发用于膜蛋白研究的无洗涤剂天然细胞膜纳米颗粒系统。这 天然细胞膜纳米颗粒系统由三个组成部分组成：1）膜活动 聚合物。 2）制备天然细胞膜纳米颗粒的方案。 3）单粒子冷冻EM 天然细胞膜纳米颗粒的分析。我们建议开发一个可以通常的系统 适用于原核生物和真核膜蛋白。特定目标1专注于施工 膜活动聚合物库的库，并开发用于制备天然细胞膜的方案 使用我们选择的模型膜蛋白的纳米颗粒。我们已经开发了四个原型膜 活性聚合物，我们计划采用类似的策略来扩展膜活性聚合物 实验室合成的市售苯乙烯马利甲基酸酐共聚物和聚合物。 特定的AMI 2专注于开发用于高分辨率结构确定所选协议 原核模型膜蛋白。特定目标3专注于为高分辨率开发协议 选定的真核模型膜蛋白的结构。拟议的研究很重要，因为它 提供了一种无革命性的清洁剂方法，用于研究膜蛋白的本地脂质 环境。正如我们在初步数据中所证明的那样，我们发现了 膜蛋白的蛋白质脂质相互作用对于理解理解的结构和功能至关重要 膜蛋白。该系统可能会改变膜蛋白研究领域，并具有很好的 对基于膜蛋白结构的药物发现和发育的影响。