Native Cell Membrane Nanoparticles System

摘要 膜蛋白在生物体中起着至关重要的作用。蛋白质-脂质的相互作用对于 膜蛋白的结构和功能完整性。膜蛋白结构的高分辨率 对了解它们的生物学和基于结构的药物发现都有很高的要求。这是经常需要的 从细胞膜中提取膜蛋白，用于生化和生物物理研究。目前 洗涤剂主要用于膜蛋白的提取。洗涤剂有很大的缺点 因为它们破坏脂双层，并从膜蛋白中去除重要的脂分子。我们是 开发用于膜蛋白研究的免洗涤剂天然细胞膜纳米颗粒系统。这个 天然细胞膜纳米粒子系统由三部分组成：1)膜活性 聚合物。2)天然细胞膜纳米粒的制备方案。3)单粒子低温电磁 天然细胞膜纳米颗粒的分析。我们建议开发一个系统，它可以普遍地 适用于原核生物和真核生物的膜蛋白。具体目标1侧重于建设 膜活性聚合物文库的建立和天然细胞膜制备方案的开发 纳米颗粒使用我们选择的模型膜蛋白。我们已经开发了四个原型膜 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 市面上可买到的苯乙烯-马来酸酐共聚物和实验室合成的聚合物。 特定的AMI2专注于开发用于所选化合物的高分辨率结构确定的协议 原核模型膜蛋白。具体目标3侧重于开发高分辨率的协议 筛选的真核模型膜蛋白的结构。这项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免洗涤剂方法 环境。正如我们在初步数据中所展示的那样，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的研究领域，具有很大的应用价值 对基于膜蛋白结构的药物发现和开发的影响。

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.