Collaborative Research: Single Cell Proteomics

合作研究：单细胞蛋白质组学

• 批准号: 0138048
• 负责人: Steven Soper
• 金额: $ 82.07万
• 依托单位: Louisiana State University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-04-01 至 2006-03-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0138048&HistoricalAwards=false
• 关键词: Collaborative; Research; Single; Cell; Proteomics

This award provides support for a joint project aimed at the development of aggressive new analytical strategies for protein sequence analysis based on mass spectrometry. The effort involves investigators at Louisiana State University, who are accomplished in the manufacture of micrometer-sized devices, and their collaborator at the University of Cincinnati, who is skilled in high resolution mass spectrometry. Microfabrication techniques will be used to develop integrated micro-systems expected to permit analysis of the proteins in a single cell. The use of microfludics will help maintain the proteins at a high enough concentration to permit analysis even though the amount of each protein found in a cell is on the order of 3000 molecules. The devices to be used in this project are built using polymers and micromanufacturing techniques to fabricate high-aspect-ratio molding dies for hot embossing polymers. The devices will be tested and refined through studies of chloroplast-derived membranes thought to contain at least 150 different proteins. Within the context of the proposed effort, these studies will be extended to examine the proteome, i.e., the entire complement of proteins, of intact chloroplasts of Arabadopsis and of entire cells of the blue-green algae Synechocystis. If fully successful, the instrumentation will allow biochemical analyses of individual cells and thus contribute to a better understanding of the relationship of gene expression and cellular phenotype.

该奖项为旨在开发基于质谱的蛋白质序列分析的积极新分析策略的联合项目提供支持。 这项工作涉及路易斯安那州州立大学的研究人员，他们在制造微米尺寸的设备方面很有成就，他们在辛辛那提大学的合作者，擅长高分辨率质谱分析。 微加工技术将被用来开发集成的微系统，预计允许在一个单一的细胞中的蛋白质的分析。 微流体的使用将有助于将蛋白质保持在足够高的浓度以允许分析，即使在细胞中发现的每种蛋白质的量在3000个分子的数量级上。 在这个项目中使用的设备是建立使用聚合物和微制造技术，以制造高纵横比的热压聚合物成型模具。 这些设备将通过研究叶绿体衍生的膜进行测试和改进，这些膜被认为含有至少150种不同的蛋白质。在拟议的努力范围内，这些研究将扩展到检查蛋白质组，即，蛋白质的完整补充，阿拉伯藻的完整叶绿体和蓝绿藻集胞藻的整个细胞。 如果完全成功，该仪器将允许对单个细胞进行生化分析，从而有助于更好地理解基因表达和细胞表型的关系。