Proposal for an Administrative Supplement for Equipment Purchases for NIGMS Awardees NOT-GM-22-017

父拨款的项目摘要R01GM132329 膜蛋白在生物体中是必不可少的。蛋白质-脂质相互作用是膜的关键 蛋白质结构和功能的完整性。高分辨率膜蛋白结构处于高水平 需要了解它们的生物学和基于结构的药物开发。提取 生物化学和生物物理研究常常需要从细胞膜中提取膜蛋白。 目前，洗涤剂是提取膜蛋白最常用的方法。洗涤剂 有几个缺点，包括破坏脂双层和去除必要的脂类 来自膜蛋白的分子。我们正在创造一种不含洗涤剂的天然细胞膜 纳米技术在膜蛋白研究中的应用。天然细胞膜纳米粒 体系由三部分组成：1)膜活性聚合物。2)创建原生文件的方法 细胞膜纳米颗粒。3)天然细胞膜的低温电子显微镜单颗粒研究 纳米粒子。我们建议建立一种既适用于细菌膜又适用于真核细胞膜的系统 蛋白质。具体目标1专注于构建制造天然细胞膜纳米颗粒的技术 利用我们选择的模型膜蛋白，构建了膜活性聚合物文库。我们 已经创造了四种膜活性聚合物的原型；我们打算扩大活性膜 使用商业上可获得的苯乙烯-马来酸酐共聚物的聚合物和制造的聚合物 在使用类似程序的实验室中。特定的AMI 2侧重于创建程序以确定 所选细菌模型膜蛋白的高分辨结构。第三个具体目标是 为选定的真核模型膜蛋白的高分辨率结构创建程序。这个 提出的研究很有趣，因为它为研究膜提供了一种新的免洗涤剂方法。 蛋白质在其天然的脂类环境中。正如我们的第一个数据显示的那样，蛋白质-脂质的结构知识 膜蛋白之间的相互作用是理解膜结构和功能的关键 蛋白质。这种方法有可能彻底改变膜蛋白研究领域和 对基于膜蛋白结构的药物的发现和发展具有重大影响。

Project Summary for Parent Grant R01GM132329 Membrane proteins are essential in living organisms. Protein-lipid interactions are critical for membrane protein structural and functional integrity. High-resolution membrane protein structures are in high demand for understanding their biology and drug development based on structure. Extraction of membrane proteins from cell membranes is frequently required for biochemical and biophysical research. Currently, detergents are the most often used method for extracting membrane proteins. Detergents have several downsides, including the destruction of lipid bilayers and the removal of essential lipid molecules from membrane proteins. We are creating a detergent-free native cell membrane nanoparticles technology for membrane protein research. The native cell membrane nanoparticles system is made up of three parts: 1) Polymers that are membrane-active. 2) Methods for creating native cell membrane nanoparticles. 3) Cryo-EM single particle investigation of native cell membrane nanoparticles. We suggest creating a system that is applicable to both bacterial and eukaryotic membrane proteins. Specific Aim 1 focuses on building techniques to create native cell membrane nanoparticles utilizing our chosen model membrane proteins and constructing a membrane-active polymer library. We have created four prototype membrane active polymers; we intend to expand the active membrane polymers utilizing both commercially available styrene-maleic anhydride copolymers and polymers made in laboratories using similar procedures. Specific Ami 2 focuses on creating procedures for determining the high-resolution structure of selected bacterial model membrane proteins. The third specific aim is to create procedures for high-resolution structures of selected eukaryotic model membrane proteins. The proposed study is interesting because it offers a novel detergent-free method for investigating membrane proteins in their native lipid context. As our first data show, structural knowledge of protein-lipid interactions for membrane proteins is critical for understanding the structure and function of membrane proteins. This approach has the potential to revolutionize the area of membrane protein research and have a significant impact on membrane protein structure-based medication discovery and development.