CAREER: Global analysis of in vivo protein folding efficiencies by mass spectrometry

蛋白质最初合成为线性聚合物，随后必须折叠成精确的三维结构。在细胞内，这种折叠反应往往容易出错，并可能导致错误折叠和无功能的蛋白质。该细胞包含许多质量控制机制，用于降解错误折叠和无功能蛋白质。在这个项目中，加姆马加米博士？他的实验室将开发一种新的技术，用于测量合成后在细胞内成功折叠的蛋白质的分数。利用这项技术，Ghaemmaghami博士将研究几种蛋白质折叠效率的变化。这些结果将阐明蛋白质的物理性质与其在细胞内成功折叠的倾向之间的关系。总的来说，这项研究将建立新的技术，这将有助于研究细胞内的蛋白质折叠，并导致更好地了解影响蛋白质稳态的因素。该提案的综合教育活动包括创建一个互动的、基于网络的教学模块，向学生和公众介绍现代蛋白质组学的理论和实践方面。此外，还将为高中和本科学生提供一些研究机会。该项目的多学科性质（包括蛋白质生物化学，细胞生物学，分析化学，物理化学和定量生物学）将为研究生提供跨部门的研究机会。这项研究将导致质谱相关协议和新的分析软件的发展，并将促进罗切斯特大学的蛋白质组学研究。它还将为跨部门合作研究提供基础。该软件将免费向研究界提供，并可供对动态蛋白质组学感兴趣的研究人员广泛使用。

Proteins are initially synthesized as linear polymers and must subsequently fold into precise three-dimensional structures. Within a cell, this folding reaction is often error prone and can lead to misfolded and nonfunctional proteins. The cell contains a number of quality control mechanisms for degrading misfolded and nonfunctional proteins. In this project, Dr. Ghaemmaghami?s laboratory will develop a novel technique for measuring the fraction of proteins that successfully fold within the cell after synthesis. Using this technique, Dr. Ghaemmaghami will study variations in the folding efficiencies of several proteins. The results will elucidate relationships between physical properties of proteins and their propensity to fold successfully within a cell. On the whole, this research will establish novel techniques that will facilitate investigations of protein folding inside cells and lead to a greater understanding of factors that influence protein homeostasis.Broader impacts. Integrated educational activities of the proposal include creating an interactive, web-based teaching module that introduces students and the public to theoretical and practical aspects of modern proteomics. Additionally, a number of research opportunities will be provided to high school and undergraduate students. The multidisciplinary nature of the project (encompassing protein biochemistry, cell biology, analytical chemistry, physical chemistry and quantitative biology) will provide cross-departmental research opportunities to graduate students. The research will lead to the development of mass spectrometry related protocols and novel analysis software and will facilitate proteomic research at the University of Rochester. It will also provide a foundation for cross-departmental collaborative research. The software will be made freely available to the research community and could be widely used by researchers interested in dynamic proteomics.