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

NIGMS 获奖者设备采购行政补充提案 NOT-GM-22-017

基本信息

批准号：
10645526
负责人：
Youzhong Guo
金额：
$ 23.61万
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 2024-06-30
项目状态：
已结题

项目摘要

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.

家长资助项目摘要 R01GM132329 膜蛋白对于生物体至关重要。蛋白质-脂质相互作用对于膜至关重要蛋白质结构和功能的完整性。高分辨率膜蛋白结构是在高需要了解其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经常需要来自细胞膜的膜蛋白。目前，去污剂是提取膜蛋白最常用的方法。洗涤剂有几个缺点，包括破坏脂质双层和去除必需脂质来自膜蛋白的分子。我们正在创造一种不含洗涤剂的天然细胞膜用于膜蛋白研究的纳米颗粒技术。天然细胞膜纳米颗粒系统由三部分组成：1）膜活性聚合物。 2）创建native的方法细胞膜纳米颗粒。 3) 天然细胞膜的冷冻电镜单粒子研究纳米颗粒。我们建议创建一个适用于细菌和真核细胞膜的系统蛋白质。具体目标 1 侧重于构建天然细胞膜纳米粒子的技术利用我们选择的模型膜蛋白并构建膜活性聚合物库。我们创造了四种原型膜活性聚合物；我们打算扩大活性膜利用市售苯乙烯-马来酸酐共聚物和制成的聚合物的聚合物在实验室中使用类似的程序。 Specific Ami 2 侧重于创建确定程序选定细菌模型膜蛋白的高分辨率结构。第三个具体目标是创建所选真核模型膜蛋白高分辨率结构的程序。这拟议的研究很有趣，因为它提供了一种新颖的无洗涤剂方法来研究膜蛋白质在其天然脂质环境中。正如我们的第一个数据所示，蛋白质-脂质的结构知识膜蛋白的相互作用对于理解膜的结构和功能至关重要蛋白质。这种方法有可能彻底改变膜蛋白研究领域对基于膜蛋白结构的药物发现和开发具有重大影响。