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.

该奖项为基于质谱的蛋白质序列分析的积极新分析策略的发展提供了支持。 这项工作涉及路易斯安那州立大学的调查人员，他们在制造微米大小的设备方面及其辛辛那提大学的合作者，他们在高分辨率的质谱法中熟练。 微生物技术将用于开发预期的综合微型系统，以允许对单个细胞中的蛋白质进行分析。 微塑料的使用将有助于将蛋白质保持在足够高的浓度上，以允许分析，即使细胞中发现的每个蛋白质的量均为3000个分子。 该项目中使用的设备是使用聚合物和微制造技术制造的，以制造高光谱的比例成型模具，用于热压聚合物。 这些设备将通过研究叶绿体衍生的膜至少包含150种不同蛋白质的膜进行测试和完善。在提出的努力的背景下，将扩展这些研究以检查蛋白质组，即蛋白质的全部补体，阿拉伯蛋白酶完整的叶绿体和蓝绿色藻类藻类藻类的整个细胞的蛋白质。 如果完全成功，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与细胞表型的关系。