Identification of Anandamide Transport Proteins

Anandamide 转运蛋白的鉴定

• 批准号: 6859315
• 负责人: ERIC L BARKER
• 金额: $ 14.91万
• 依托单位: PURDUE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-30 至 2006-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6859315
• 关键词: SDS polyacrylamide gel electrophoresis; affinity labeling; anandamide; caveolas; cell line; cell membrane; chemical synthesis; intracellular transport; mass spectrometry; polymerase chain reaction; protein binding; protein quantitation /detection; protein structure function; proteomics; tissue /cell culture; transport proteins; western blottings

DESCRIPTION (provided by applicant): The broad, long-term objective of this R21 CEBRA application is to advance the overall understanding of molecular mechanisms regulating endogenous cannabinoid signaling by identifying proteins responsible for endocannabinoid uptake. The endogenous cannabinoids such as anandamide are fatty acid-derived molecules whose metabolism and signal termination appear to rely upon transport back into releasing cells for subsequent breakdown by an intracellular amidohydrolase enzyme. Thus, the uptake process is thought to play a critical role in the dynamic regulation of endogenous cannabimimetic activity. The Specific Aim of this proposal is to identify proteins involved with the uptake and intracellular transport of the endocannabinoid anandamide. This project has direct health-relatedness as the endogenous cannabinoid system has been implicated in modulating various physiological and pathological processes including mood, anxiety, psychosis, motor movement, inflammation, blood pressure, and pain. Recent evidence suggests that "lipid raft-" or caveolae-related endocytosis may be involved with internalization and cellular transport of anandamide. Thus, a carrier protein for anandamide would be predicted to reside in this membrane microdomain. By developing novel probes and using cutting-edge proteomic methods, putative anandamide binding proteins will be identified. The role of these proteins in anandamide uptake will be explored using modern molecular, biochemical, and pharmacological approaches. These studies will provide a basis for future studies examining the molecular mechanisms associated with both structure/function and regulation of endogenous cannabinoid transport.

描述（由申请人提供）： 这种R21 CEBRA应用的长期长期目标是通过鉴定负责内源性大麻素摄取的蛋白质来提高调节内源性大麻素信号传导的总体理解。内源性大麻素（例如蜘蛛胺）是脂肪酸衍生的分子，其代谢和信号终止似乎依赖于转运恢复到释放细胞中，以便通过细胞内氨基水性水解酶破坏。因此，摄取过程被认为在内源性大麻活性的动态调节中起关键作用。该提案的具体目的是鉴定与内源性大麻蛋白武吻合的摄取和细胞内转运有关的蛋白质。该项目具有直接与健康相关性，因为内源性大麻素系统与调节各种生理和病理过程有关，包括情绪，焦虑，精神病，运动，运动，炎症，血压和疼痛。最近的证据表明，“脂质筏 - ”或与小窝相关的内吞作用可能与anandamide的内在化和细胞转运有关。因此，将预测用于anandamide的载体蛋白驻留在该膜微域中。通过开发新型探针并使用尖端的蛋白质组学方法，将确定推定的肛门酰胺结合蛋白。这些蛋白质在Anandamide摄取中的作用将使用现代分子，生化和药理学方法探索。这些研究将为未来研究的基础研究与内源性大麻素转运的结构/功能和调节相关的分子机制。