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）： R21 CEBRA 应用的广泛、长期目标是通过鉴定负责内源性大麻素摄取的蛋白质，促进对调节内源性大麻素信号转导的分子机制的整体理解。内源性大麻素（例如 anandamide）是脂肪酸衍生的分子，其代谢和信号终止似乎依赖于转运回释放细胞，随后被细胞内酰胺水解酶分解。因此，摄取过程被认为在内源性大麻模拟活性的动态调节中发挥着关键作用。该提案的具体目的是鉴定与内源性大麻素 anandamide 的摄取和细胞内转运有关的蛋白质。该项目与健康有直接关系，因为内源性大麻素系统参与调节各种生理和病理过程，包括情绪、焦虑、精神病、运动、炎症、血压和疼痛。最近的证据表明，“脂筏”或小凹相关的内吞作用可能与 anandamide 的内化和细胞转运有关。因此，预计 anandamide 的载体蛋白将驻留在该膜微域中。通过开发新型探针并使用尖端蛋白质组学方法，将鉴定推定的 anandamide 结合蛋白。将使用现代分子、生化和药理学方法探索这些蛋白质在 anandamide 摄取中的作用。这些研究将为未来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奠定基础。