Identification of Anandamide Transport Proteins

描述（由申请人提供）： 这项R21 CEBRA应用的广泛，长期目标是通过识别负责内源性大麻素摄取的蛋白质来促进对调节内源性大麻素信号传导的分子机制的全面理解。内源性大麻素如大麻素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拟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参与内源性大麻素anandamide的摄取和细胞内转运的蛋白质。该项目与健康直接相关，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”或小窝相关的内吞作用可能参与花生四烯酸的内化和细胞转运。因此，预测anandamide的载体蛋白将驻留在该膜微区中。通过开发新的探针和使用尖端的蛋白质组学方法，将确定推定的大麻素结合蛋白。这些蛋白质在花生四烯酸吸收中的作用将使用现代分子，生物化学和药理学方法进行探索。这些研究将为未来研究与内源性大麻素转运的结构/功能和调控相关的分子机制提供基础。

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.