4D functional mapping of glucose metabolism in Living Cells

项目总结/摘要 本提案的目的是阐明葡萄糖代谢（即糖酵解和糖异生） 和线粒体代谢在空间和功能上（空间功能上）相互关联， 在人类活细胞中精心安排。目前对高度交织的代谢途径网络的理解 在2D中受到限制，但细胞代谢发生在空间和时间（即4D）中。许多代谢 途径在空间上局限于膜结合的细胞器或无膜区室。到 了解代谢途径是如何调节和网络化的，了解细胞器或 无膜区室在空间上排列并在4D中功能性地相互作用。我们最近 报道，细胞质中，葡萄糖代谢中的速率决定酶在空间上组织成 在人类细胞中有各种大小的无膜区室。我们提出它们将葡萄糖流分流到 合成代谢途径更重要的是，我们的初步结果表明， 通过液-液相分离（LLPS）形成的隔室可能在空间上和功能上与 线粒体因此，在这个提议中，我们假设葡萄糖代谢中的酶区室 在空间和功能上与线粒体在4D，LLSP发挥作用，以适应细胞 要求。我们将描述酶的形成和调节的精确机制， 隔间此外，我们将揭示葡萄糖代谢的酶室是如何 在功能上和空间上与线粒体及其网络的调节机制协调， 活细胞拟议的工作将提供新的，强大的4D成像和分析方法，以探索新的 细胞内代谢网络的时空动力学观点。这项工作将提供 理解细胞中新的基本细胞器类别的基本原则，并将提供一个新的 范例来理解活细胞中代谢网络的4-D图。

Project Summary/Abstract The objective of this proposal is to elucidate how glucose metabolism (i.e. glycolysis and gluconeogenesis) and mitochondrial metabolism are spatially and functionally (spatiofunctionally) interconnected and dynamically orchestrated in human living cells. Current understanding of the highly enmeshed web of metabolic pathways has been limited in 2D, but cellular metabolism takes place in space and time (i.e. 4D). A number of metabolic pathways are spatially confined into either membrane-bound organelles or membraneless compartments. To understand how metabolic pathways are regulated and networked, it is vital to know how organelles or membraneless compartments are spatially arranged and functionally interplay in 4D. We have recently reported that the cytoplasmic, rate-determining enzymes in glucose metabolism are spatially organized into membraneless compartments in various sizes in human cells. We proposed that they shunt glucose flux to anabolic biosynthetic pathways. More importantly, our preliminary results suggest now that the enzyme compartments, formed by liquid-liquid phase separation (LLPS), might be spatially and functionally linked with mitochondria. Thus, in this proposal, we hypothesize that the enzyme compartments in glucose metabolism are spatially and functionally associated with mitochondria in 4D, by which LLSP plays a role to adapt cellular demands. .We will characterize precise mechanisms of the formation and modulation of the enzyme compartments. In addition, we will reveal how the enzyme compartments of glucose metabolism are functionally and spatially coordinated with the mitochondria and the regulatory mechanisms of their network in living cells. The proposed work will offer new, powerful 4D imaging and analysis approaches to explore novel perspectives of spatiotemporal dynamics of metabolic networks inside cells. This work will provide the fundamental principle of understanding the new class of essential organelles in the cell and will provide a new paradigm to comprehend 4-D map of metabolic networks in living cells.