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.

摘要 信号代谢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内的 稳态和整合代谢和转录反应。代谢物转运缺陷， 分布可能导致多种疾病，包括癌症，免疫，炎症和代谢 紊乱亚细胞区室化允许相同的分子参与不同的生物学过程。 流程.信号代谢物通常作为特定蛋白质的第二信使或传感器的配体 和核受体（NR），配体激活的转录因子，感知环境信号， 细胞反应。由于其固有的反应性，NR配体的细胞内水平，沿着其 亚细胞定位受到严格控制，并可能根据营养状态发生很大的波动， 病理生理条件。尽管我们了解它们的功能，但我们对核如何 受体配体穿过细胞器的旅行仍然有限，这是由于缺乏靶向这种受体配体的特异性工具。 机制等我们建议整合化学蛋白质组学、代谢组学和细胞分析，开发新的 化学工具来询问NR配体的蛋白质相互作用组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 的信号代谢物，并确定专用的蛋白伴侣，结合NR配体在其合成位点 或进入细胞，将它们运输到细胞核，并将它们递送到核受体。我们初步调查的一个重要发现 研究发现PGRMC 2是一种细胞内血红素伴侣，可将血红素从线粒体中转运出来 并调节血红素应答转录因子如Rev-1的转录活性。 Erb β和BACH 1。我们将利用从这一初步工作中获得的经验， 已知NR配体如激活PPARs的脂肪酸的其它转运机制的鉴定， 调节代谢和系统能量稳态的配体激活转录因子家族。的 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谱，我们 期望超越细胞内贩运机制的识别，并对细胞内贩运机制产生更广泛的影响。 领域提供了一个强大的战略，研究代谢产物蛋白质串扰。最后，这个项目独特地结合了 我们在转录调控、代谢和化学生物学方面的多学科专业知识， 探索细胞生物学中令人兴奋的新发现。