Collaborative Research: Development of a platform enabling analysis of membrane protein interactions

合作研究：开发能够分析膜蛋白相互作用的平台

• 批准号: 1615252
• 负责人: Barbara Imperiali
• 金额: $ 15万
• 依托单位: Massachusetts Institute of Technology
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-08-01 至 2019-06-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1615252&HistoricalAwards=false
• 关键词: Collaborative; Research; Development; platform; enabling

Title: Collaborative research: Development of a platform enabling analysis of membrane protein interactionsProteins embedded within or associated with cellular membranes make up 30% of all proteins. These cellular gatekeepers are critical in biological functions, including transport in and out of cells, recognition and transduction of information, energy production and communication amongst cells. Despite the importance of membrane proteins, understanding of the interactions and functions of the membrane proteome is still greatly hampered by technical challenges; there is an urgent need for innovative approaches to uncover how membrane proteins interact with one another and with their binding partners in the native membrane environment. This project will develop a versatile experimental platform that provides maps of protein-protein and protein-membrane interactions. Under-represented minorities will receive STEM education and training at undergraduate graduate level. The proposal will also provide video instructions for high school students to attract students in STEM fields. This collaborative research will establish a versatile experimental platform that provides information on protein-protein and protein-membrane assemblies in Nanodiscs, which represent a native-like model membrane system. The goals of this research are to establish a versatile experimental platform that provides vector maps of protein-protein and protein-membrane assemblies. Key cornerstones of the approach are the use of lipid bilayer nanodiscs (NDs) as a native-like membrane platform and biophysical approaches that rely on strategically positioned lanthanide-binding tags (LBTs). The LBTs are dual-function probes that insert atoms for physical measurements enabling the construction of three-dimensional vector maps of protein assemblies relative to the membrane plane. Measurements will be accomplished via small-angle X-ray scattering (SAXS) and anomalous SAXS (ASAXS) to provide distances between lanthanides and center of mass and inter-lanthanide distances. The construction of vector maps will allow accurate measurement of the response of proteins to the presence and absence of binding partners, dynamic protein modifications, and indeed the composition of the membrane itself-thus defining the critical determinants of both assembly and function. This project is supported by the Molecular Biophysics Cluster of the Molecular and Cellular Biosciences Division in the Directorate for Biological Sciences.

标题：合作研究：开发一个能够分析膜蛋白相互作用的平台嵌入细胞膜中或与细胞膜相关的蛋白质占所有蛋白质的30%。这些细胞守门人在生物学功能中至关重要，包括细胞内和细胞外的运输、信息的识别和转导、能量生产和细胞间的通信。尽管膜蛋白很重要，但膜蛋白质组的相互作用和功能的理解仍然受到技术挑战的极大阻碍；迫切需要创新的方法来揭示膜蛋白在天然膜环境中如何相互作用及其与结合伙伴的作用。该项目将开发一个多功能的实验平台，提供蛋白质-蛋白质和蛋白质-膜相互作用的地图。代表不足的少数群体将接受本科生研究生水平的STEM教育和培训。该提案还将为高中生提供视频指导，以吸引STEM领域的学生。这项合作研究将建立一个通用的实验平台，提供关于纳米盘中蛋白质-蛋白质和蛋白质-膜组装的信息，这代表了一个类似于原生的模型膜系统。这项研究的目标是建立一个通用的实验平台，提供蛋白质-蛋白质和蛋白质-膜组装的矢量图。该方法的主要基石是使用脂质双层纳米盘(NDS)作为天然的膜平台，以及依赖于战略性定位的稀土结合标签(LBT)的生物物理方法。LBTS是双功能探测器，可以插入原子进行物理测量，从而能够构建相对于膜平面的蛋白质组装的三维矢量图。测量将通过小角X射线散射(SAXS)和反常SAXS(ASAXS)来完成，以提供稀土元素与质心之间的距离和稀土元素间的距离。载体图的构建将允许准确测量蛋白质对结合伙伴的存在和不存在的响应，动态的蛋白质修饰，甚至膜本身的组成-从而确定组装和功能的关键决定因素。该项目得到了生物科学局分子和细胞生物科学司分子生物物理组的支持。