Off the beaten path(way): Spatiotemporal investigation of protein assemblies controlling mitochondrial metabolism

不走寻常路：控制线粒体代谢的蛋白质组装体的时空研究

• 批准号: 10244772
• 负责人: Breann Brown
• 金额: $ 131.32万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-21 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10244772
• 关键词: 3-Dimensional; Address; Area; Biochemical Pathway; Biology; Breathing; Communities; Complex; Crowding; Cryoelectron Microscopy; Defect; Diabetes Mellitus; Environment; Erythrocytes; Heme; Human; Immune response; Inflammation; Investigation; Knowledge; Life; Lipids; Metabolic; Metabolic Diseases; Metabolism; Output; Pathway interactions; Plants; Process; Production; Proteins; Proteomics; Research; Resolution; Signal Transduction; Structure; System; Textbooks; Time; Vacuum; anticancer research; biological systems; cofactor; experimental study; insight; macromolecule; metabolomics; mitochondrial metabolism; movie; oxygen transport; small molecule; spatiotemporal; tumor progression

Cellular metabolism is the crux of all organismal biology. Therefore, uncovering fundamental knowledge regarding how metabolism is controlled will have far-reaching implications. Metabolic systems are traditionally depicted as linear or circular pathways in textbooks. In reality, these processes are intricately governed by complex, higher-order networks of macromolecules including proteins and lipids. A metabolon is a dynamic cluster of proteins, cofactors, and small molecules that interact to control a metabolic process. Importantly, metabolons are found across multiple biological systems from plants to humans, indicating their fundamental importance in biology. Heme is an essential and conserved biomolecule that is produced by the community of proteins forming the heme metabolon. Heme not only transports oxygen in red blood cells, but it also serves as a catalytic cofactor for proteins governing multiple cellular signaling processes across all kingdoms of life. Thus, determining how proteins assemble and disassemble to control heme metabolon formation will provide insight into production of this critical molecule and also form the basis for studying other key metabolons. Specifically, we will 1) isolate and solve the structure of the heme metabolon, 2) determine dynamics of metabolon formation, and 3) investigate how defects in specific assembly steps alter metabolic output. We will accomplish this by integrating high-resolution cryo-EM with time-resolved proteomics and metabolomics experiments to reveal metabolon dynamics. The combination of these approaches will unite multiple hierarchies of cellular signaling, transforming the static textbook snapshot of metabolism into a 3D movie of a living, breathing metabolic machine. Addressing the fundamental and unknown question of how metabolic networks are controlled via coordinated protein organization will have major impacts in broad areas of research, including cancer progression, diabetes, and the immune response.

细胞新陈代谢是所有生物生物学的关键。因此，揭开基础知识 关于如何控制新陈代谢将具有深远的影响。新陈代谢系统传统上 在教科书中被描绘成直线或圆形的小路。在现实中，这些过程由错综复杂的 包括蛋白质和脂类在内的复杂的高阶大分子网络。新陈代谢是一种动力 蛋白质、辅因子和小分子相互作用以控制新陈代谢过程的簇。重要的是 从植物到人类的多个生物系统中都发现了代谢物，这表明了它们的基本原理 在生物学中的重要性。血红素是一种必需的和保守的生物分子，由 形成血红素代谢素的蛋白质。血红素不仅能运输红细胞中的氧气，还能起到 蛋白质的催化辅因子，控制着所有生命王国的多个细胞信号过程。因此， 确定蛋白质如何组装和分解以控制血红素代谢的形成将提供洞察力 这种关键分子的产生，也形成了研究其他关键代谢物的基础。具体来说， 我们将1)分离和解决血红素代谢物的结构，2)确定代谢物形成的动力学， 以及3)研究特定组装步骤中的缺陷如何改变新陈代谢产出。我们将通过以下方式实现这一目标 将高分辨率冷冻-EM与时间分辨蛋白质组和代谢组学实验相结合，揭示 代谢动力学。这些方法的组合将统一细胞信号的多个层次， 将教科书上关于新陈代谢的静态快照转变成一部关于活的、会呼吸的新陈代谢机器的3D电影。 解决代谢网络如何通过协调控制这一根本而未知的问题 蛋白质组织将在广泛的研究领域产生重大影响，包括癌症进展、糖尿病、 和免疫反应。