Native Cell Membrane Nanoparticles System

总结 膜蛋白在生物体中起着至关重要的作用。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 对了解它们的生物学和基于结构的药物发现都有很高的需求。往往需要 从细胞膜中提取膜蛋白用于生物化学和生物物理学研究。目前 去污剂主要用于膜蛋白提取。洗涤剂有明显缺点 因为它们破坏脂质双层并从膜蛋白中除去重要脂质分子。我们 开发用于膜蛋白研究的无去污剂天然细胞膜纳米颗粒系统。的 天然细胞膜纳米颗粒系统由三个组分组成：1）膜活性 聚合物2)天然细胞膜纳米颗粒的制备方案。3)单粒子冷冻电镜 天然细胞膜纳米颗粒的分析。我们建议开发一个系统， 适用于原核和真核细胞膜蛋白。具体目标1侧重于建设 膜活性聚合物库，并开发用于制备天然细胞膜的方案 纳米颗粒使用我们选择的模型膜蛋白。我们已经开发了四种原型膜 活性聚合物，与类似的战略，我们计划扩大膜活性聚合物使用这两个 市售苯乙烯马来酸酐共聚物和实验室合成的聚合物。 Specific Ami 2专注于开发用于选择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 选择的真核模型膜蛋白的结构。这项研究意义重大，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洗涤剂方法 环境正如我们在初步数据中所证明的那样，我们发现了 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。 膜蛋白这一系统将改变膜蛋白的研究领域， 对基于膜蛋白结构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.