Dissecting intracellular metabolite trafficking using chemoproteomics

使用化学蛋白质组学剖析细胞内代谢物运输

• 批准号: 10711934
• 负责人: Andrea Galmozzi
• 金额: $ 38.88万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-01 至 2028-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10711934
• 关键词: Automobile Driving; BACH1 gene; Binding; Biological Process; Biology; Cell Cycle; Cell Nucleus; Cell physiology; Cells; Cellular Assay; Cellular biology; Chemicals; Dedications; Defect; Disease; Family; Fatty Acids; Genetic Transcription; Goals; Heme; Homeostasis; Immunologics; Inflammatory; Intracellular Transport; Knowledge; Ligands; Malignant Neoplasms; Maps; Metabolic; Metabolic Diseases; Metabolism; Mitochondria; Molecular; Molecular Chaperones; Mus; Nuclear Receptors; Nutritional; Organelles; Peroxisome Proliferator-Activated Receptors; Physiology; Proteins; Proteome; Regulation; Second Messenger Systems; Signal Transduction; Site; Stimulus; Technology; Therapeutic Agents; Time; Transcriptional Regulation; Travel; Work; chemoproteomics; experience; metabolomics; multidisciplinary; novel; protein metabolite; receptor function; response; sensor; tool; trafficking; transcription factor

ABSTRACT Signaling metabolites control various cellular processes, including cell cycle, differentiation, and adaptations to environmental stimuli. Intracellular trafficking of signaling metabolites is crucial for maintaining cellular homeostasis and integrate metabolic and transcriptional responses. Defects in metabolite transport and distribution may lead to multiple diseases, including cancer, immunological, inflammatory, and metabolic disorders. Subcellular compartmentalization allows the same molecules to partake in distinct biological processes. Signaling metabolites generally act as second messengers for specific proteins or ligands for sensors and nuclear receptors (NR), ligand-activated transcription factors that sense environmental signals and drive cellular response. Because of their intrinsic reactivity, the intracellular levels of NR ligands, along with their subcellular localization, are tightly controlled and may oscillate greatly depending on nutritional states and pathophysiological conditions. Despite our understanding of their functions, our knowledge of how nuclear receptor ligands travel across organelles remains limited due to the lack of specific tools to target such mechanisms. We propose to integrate chemoproteomics, metabolomics, and cellular assays, to develop novel chemical tools to interrogate the protein interactomes of NR ligands and identify their intracellular chaperones. Leveraging these technologies, we intend to reveal the molecular and functional basis of intracellular trafficking of signaling metabolites and identify dedicated protein chaperones that bind NR ligands at their site of synthesis or entry into the cell, transport them to the nucleus, and deliver them to NRs. A driving finding of our preliminary work was the discovery of PGRMC2 as an intracellular heme chaperone that transports heme from mitochondria to the nucleus and regulates the transcriptional activity of heme-responsive transcription factors such as Rev- Erb and BACH1. We will use the experience acquired from this initial work to extend our studies to the identification of other transport mechanisms for known NR ligands, such as fatty acids, that activate PPARs, a family of ligand-activated transcription factors that regulate metabolism and systemic energy homeostasis. The second major goal of this proposal is to develop spatial- and time-resolved protein-metabolite maps, which we expect to go beyond the identification of intracellular trafficking mechanisms and have a broader impact on the field by providing a powerful strategy to study metabolite-protein crosstalk. Lastly, this project uniquely combines our multidisciplinary expertise in transcriptional regulation, metabolism, and chemical biology to lead the exploration of a new exciting findings in cell biology.

抽象的 信号代谢产物控制各种细胞过程，包括细胞周期，分化和适应 环境刺激。信号代谢物的细胞内运输对于维持细胞至关重要 稳态和综合代谢和转录反应。代谢物运输的缺陷和 分布可能导致多种疾病，包括癌症，免疫学，炎症和代谢 疾病。亚细胞隔室化允许相同的分子参与不同的生物学 过程。信号代谢物通常充当传感器特定蛋白质或配体的第二信使 和核接收器（NR），配体激活的转录因子，感知环境信号和驱动 细胞反应。由于其内在反应性，NR配体的细胞内水平以及它们 亚细胞定位，受到严格控制，并且可能非常振荡，具体取决于营养状态和 病理生理状况。尽管我们了解了它们的功能，但我们对核如何的了解 由于缺乏针对此类的特定工具，受体配体在整个细胞器中旅行仍然有限 机制。我们建议整合化学蛋白质组学，代谢组学和细胞测定法，以开发新颖的 化学工具来询问NR配体的蛋白质相互作用并鉴定其细胞内伴侣。 利用这些技术，我们打算揭示细胞内运输的分子和功能基础 信号代谢产物并确定在合成部位结合NR配体的专用蛋白质伴侣 或进入细胞，将它们运输到细胞核中，然后将其输送到NRS。我们初步的驾驶发现 工作是发现PGRMC2是一种细胞内血红素伴侣，可从线粒体转运血红素 对核并调节血红素反应性转录因子（例如Rev-）的转录活性 Erb和Bach1。我们将利用从这项初始工作中获得的经验将我们的研究扩展到 鉴定已知NR配体（例如脂肪酸）的其他转运机制激活PPAR，A 调节代谢和全身能量稳态的配体激活转录因子家族。这 该提案的第二个主要目标是开发空间和时间分辨的蛋白质代谢物图，我们是我们的 期望超越细胞内贩运机制的识别，并对 通过提供强大的策略来研究代谢产物蛋白串扰。最后，这个项目独特地结合了 我们在转录调节，代谢和化学生物学方面的多学科专业知识，以领导 探索细胞生物学中新的令人兴奋的发现。