4D functional mapping of glucose metabolism in Living Cells

项目摘要/摘要 这项建议的目的是阐明葡萄糖代谢(即糖酵解和糖异生)是如何 线粒体代谢在空间上和功能上(空间功能上)是相互关联和动态的 在人类活细胞中精心编排。目前对高度网状的代谢途径网络的理解 仅限于2D，但细胞新陈代谢在空间和时间上发生(即4D)。一些新陈代谢 通路在空间上被限制在膜结合的细胞器或无膜的隔室中。至 了解代谢途径是如何调节和联网的，了解细胞器或 无膜隔间在4D中空间布置并在功能上相互作用。我们最近做了 报道称，葡萄糖代谢中的细胞质、速率决定酶在空间上被组织成 人体细胞中大小不一的无膜室。我们建议它们将葡萄糖流量分流到 合成代谢生物合成途径。更重要的是，我们的初步结果表明，现在这种酶 由液-液相分离(LLP)形成的隔室可能在空间和功能上与 线粒体。因此，在这个提议中，我们假设这种酶在葡萄糖代谢中起到隔间作用。 LLSP在空间上和功能上都与4D中的线粒体相关，LLSP通过线粒体发挥适应细胞的作用 要求。我们将描述这种酶的形成和调节的精确机制 车厢。此外，我们还将揭示葡萄糖新陈代谢的酶是如何 与线粒体及其网络的调控机制在功能和空间上的协调 活细胞。这项拟议的工作将为探索小说提供新的、强大的4D成像和分析方法 细胞内代谢网络的时空动力学前景。这项工作将提供 理解细胞中新的一类基本细胞器的基本原理，并将提供一种新的 理解活细胞中代谢网络的四维图的范例。

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.