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，该奖项为高度创新或概念上创造性的研究提供支持，以推进了解药物滥用和成瘾。 准确评估膜蛋白的脂质嵌入残基对于预测膜蛋白功能和生成结构模型至关重要。目前，膜蛋白拓扑预测算法最多精确到 +/- 5 到 10 个氨基酸。我们建议开发一种鸟枪式蛋白质组学方法，用于对复杂生物样品中整合膜蛋白的脂质嵌入残基进行整体分析。我们将应用这些方法来：1）一种定向方法来表征人类多巴胺转运蛋白（hDAT）的拓扑结构、hDAT 的运输突变体和大鼠脑组织中的天然 DAT；2）一种非定向的全局方法来表征膜的拓扑结构从细胞和组织匀浆中富集的蛋白质。