Subcellular distribution, trafficking and signal transduction of G protein-coupled receptors for energy metabolites

能量代谢物 G 蛋白偶联受体的亚细胞分布、运输和信号转导

• 批准号: 407707190
• 负责人: Privatdozentin Dr. Claudia Stäubert
• 金额: --
• 依托单位: Rudolf-Schönheimer-Institut für Biochemie
• 依托单位国家: 德国
• 项目类别: Research Grants
• 资助国家: 德国
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/407707190?language=en
• 关键词: Subcellular; distribution; trafficking; signal; transduction

G protein-coupled receptors (GPCRs) are the largest superfamily of transmembrane receptors but accumulating evidence reveals that GPCRs are present and functional not only at the plasma membrane but may also signal from intracellular compartments. GPCR signaling from inside the cell is an aspect highly relevant in the context of metabolite-sensing GPCRs (msGPCRs), which are activated by (intracellularly synthesized) metabolic intermediates of central processes such as glycolysis, citric acid cycle or β-oxidation. Here, we will continue our effort to better understand, if and how selected msGPCRs that reside in intracellular compartments mediate their signal from there. As one representative msGPCR, succinate receptor is in the focus, for which we aim to understand the mechanisms behind its activation, signaling and trafficking dependent on its localization. Succinate receptor exhibits a strong link to metabolism and its agonist succinate gains increasingly relevance as oncometabolite. Therefore, we intent to develop a sensor that enables detection of succinate in live cells. On the other hand, we extent our focus to the oxoeicosanoid receptor and its evolutionary closely related family of hydroxycarboxylic acid receptors (HCAR). The oxoeicosanoid receptor exhibits unique structural features, an unusual subcellular distribution and is rather poorly understood. We aim to dismantle components involved in signaling and trafficking of this receptor in comparison to the msGPCRs of the HCAR family. Using bioluminescence and fluorescence resonance energy transfer (BRET/FRET), dynamic mass redistribution and multi-parametric surface plasmon resonance analyses in combination with several signal transduction assay systems and imaging techniques, we will address the challenging question of location-dependent signal transduction and trafficking of these msGPCRs. Our results will fundamentally increase our knowledge about selected GPCRs activated by energy metabolites and provide further evidence for their ‘not plasma membrane bound’ function.Ultimately, our study will result in an extended understanding of the molecular mechanism and structural requirements of (localization-dependent) signal transduction, trafficking and pharmacology of the selected msGPCRs. These results will be of great value for the assessment of their potential as targets for the development of anti-inflammatory or anti-cancer drugs.

G蛋白偶联受体（GPCR）是最大的跨膜受体超家族，但越来越多的证据表明，GPCR不仅存在于质膜上，而且还可能从细胞内隔室发出信号。来自细胞内部的GPCR信号传导是在代谢物感应GPCR（msGPCR）的背景下高度相关的方面，msGPCR由诸如糖酵解、柠檬酸循环或β-氧化的中枢过程的（细胞内合成的）代谢中间体激活。在这里，我们将继续努力更好地了解，如果以及如何选择msGPCR驻留在细胞内隔室介导他们的信号从那里。琥珀酸受体作为msGPCR的代表性受体之一，是研究的重点，我们的目标是了解其激活、信号传导和运输依赖于其定位的机制。琥珀酸受体与代谢有密切联系，其激动剂琥珀酸作为肿瘤代谢产物的相关性日益增强。因此，我们打算开发一种能够检测活细胞中琥珀酸的传感器。另一方面，我们将我们的重点扩展到oxoeicosanoid受体及其进化密切相关的羟基羧酸受体家族（HCAR）。oxoeicosanoid受体具有独特的结构特征，不寻常的亚细胞分布，而知之甚少。我们的目标是拆除组件参与信号和运输的这种受体相比，msGPCR的HCAR家族。使用生物发光和荧光共振能量转移（BRET/FRET），动态质量再分布和多参数表面等离子体共振分析结合几个信号转导检测系统和成像技术，我们将解决这些msGPCR的位置依赖性信号转导和贩运的挑战性问题。我们的研究结果将从根本上增加我们的知识，选定的GPCR激活的能量代谢产物，并提供进一步的证据，为他们的'不质膜结合'的功能。最终，我们的研究将导致扩展的分子机制和结构要求（定位依赖）的信号转导，运输和药理学的选定msGPCR的理解。这些结果对于评估它们作为抗炎或抗癌药物开发靶点的潜力具有重要价值。