Identification of Anandamide Transport Proteins

描述(由申请人提供)： R21 CEBRA应用的广泛、长期的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控内源性大麻素信号的分子机制的整体理解。内源性大麻素，如阿南达胺，是脂肪酸衍生的分子，其新陈代谢和信号终止似乎依赖于运输回释放细胞，随后被细胞内的氨基水解酶分解。因此，摄取过程被认为在内源性大麻仿制活性的动态调节中发挥关键作用。这项建议的具体目的是确定与内源性大麻素脱氢酶的摄取和细胞内转运有关的蛋白质。这个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”或小凹相关的内吞作用可能与阿南达胺的内化和细胞转运有关。因此，阿南达胺的载体蛋白将被预测存在于这个膜微域中。通过开发新的探针和使用尖端的蛋白质组学方法，假定的anandamide结合蛋白将被识别出来。将使用现代分子、生物化学和药理学方法探索这些蛋白质在anandamide吸收中的作用。这些研究将为今后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.