Native Cell Membrane Nanoparticles System

天然细胞膜纳米粒子系统

基本信息

批准号：
10200844
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
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-07-01 至 2023-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10200844
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

项目摘要

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发挥着至关重要的作用。蛋白质-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对了解其生物学和基于结构的药物发现的需求很高。往往是需要从细胞膜中提取膜蛋白用于生化和生物物理研究。现在去污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部分组成：1）膜活性聚合物。 2）天然细胞膜纳米颗粒的制备方案。 3) 单颗粒冷冻电镜天然细胞膜纳米粒子的分析。我们建议开发一个可以通用的系统适用于原核和真核膜蛋白。具体目标 1 侧重于建设膜活性聚合物库的构建并开发天然细胞膜的制备方案使用我们选择的模型膜蛋白的纳米颗粒。我们开发了四种原型膜活性聚合物，采用类似的策略，我们计划使用这两种方法来扩展膜活性聚合物市售的苯乙烯马来酸酐共聚物和实验室合成的聚合物。 Specific Ami 2 专注于开发用于选定的高分辨率结构测定的协议原核模型膜蛋白。具体目标 3 侧重于开发高分辨率协议选定的真核模型膜蛋白的结构。拟议的研究意义重大，因为它为研究局部脂质中的膜蛋白提供了一种革命性的无去污剂方法环境。正如我们在初步数据中所证明的那样，我们发现了结构信息膜蛋白的蛋白质-脂质相互作用对于理解膜蛋白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，并具有巨大的应用前景。对基于膜蛋白结构的药物发现和开发的影响。