Proteomic Tools for the Comprehensive Analysis of Dopamine Transporter Topology

用于多巴胺转运蛋白拓扑综合分析的蛋白质组学工具

• 批准号: 7135141
• 负责人: Christine C Wu
• 金额: $ 15.24万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-05-01 至 2008-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7135141
• 关键词: dopamine transporter; membrane proteins; proteomics; tissues

DESCRIPTION (provided by applicant): This R21 application (10 pages max) is being submitted in response to PAR-03-017 "Cutting-Edge Basic Research Awards (CEBRA)" which provides support for highly innovative or conceptually creative research to advance the understanding of drug abuse and addiction. Accurate assessments of the lipid embedded residues of membrane proteins are crucial for the prediction of membrane protein function and the generation of structural models. Currently, membrane protein topology prediction algorithms are accurate to +/- 5 to 10 amino acids at best. We propose to develop a shotgun proteomic approach for the global analysis of lipid embedded residues of integral membrane proteins within a complex biological sample. We will apply these methods to: 1) a directed approach to characterize the topology of the human dopamine transporter (hDAT), trafficking mutants of hDAT, and native DAT in rat brain tissue and 2) an undirected global approach to characterize the topology of membrane proteins enriched from cell and tissue homogenates.

描述（由申请人提供）：此R21申请（最多10页）是为了响应PAR-03-017“尖端基础研究奖（CEBRA）”而提交的，该奖项为高度创新或概念创造性的研究提供支持，以促进对药物滥用和成瘾的理解。 膜蛋白脂质包埋残基的准确评估是至关重要的膜蛋白功能的预测和结构模型的生成。目前，膜蛋白拓扑预测算法最多精确到+/-5至10个氨基酸。我们建议开发一种鸟枪蛋白质组学方法，用于复杂生物样品中完整膜蛋白的脂质嵌入残基的全球分析。我们将这些方法应用于：1）一个定向的方法来表征拓扑结构的人多巴胺转运蛋白（hDAT），运输突变体的hDAT，和天然DAT在大鼠脑组织和2）一个非定向的全球性的方法来表征的拓扑结构的膜蛋白富集从细胞和组织匀浆。