Abstract
Fluorescence live-cell imaging that has contributed to our understanding of cell biology is now at the frontline of studying quantitative biochemistry in a cell. Particularly, technological advancements of fluorescence live-cell imaging and associated strategies in recent years have allowed us to discover various subcellular macromolecular assemblies in living human cells. Here we describe how real-time dynamics of a multienzyme metabolic assembly, the “glucosome,” that is responsible for regulating glucose flux at subcellular levels, has been investigated in both 2- and 3-dimensional space of single human cells. We envision that such multi-dimensional fluorescence live-cell imaging will continue to revolutionize our understanding of how intracellular metabolic pathways and their network are functionally orchestrated at single-cell levels.
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Acknowledgement
We wish to thank all former and current members who have contributed to the described protocol. We would also like to thank Dr. Gerald M. Wilson and Dr. Jiayuh Lin for sharing MCF-7 and MDA-MB-436 cell lines, respectively. This work is financially supported by the National Institutes of Health: R01GM134086 (M.K.), R01GM125981 (S.A.), R03CA219609 (S.A.), T32GM066706 (E.L.K.), and R25GM55036 (E.L.K.).
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An, S., Parajuli, P., Kennedy, E.L., Kyoung, M. (2022). Multi-dimensional Fluorescence Live-Cell Imaging for Glucosome Dynamics in Living Human Cells. In: Stamatis, H. (eds) Multienzymatic Assemblies. Methods in Molecular Biology, vol 2487. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2269-8_2
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DOI: https://doi.org/10.1007/978-1-0716-2269-8_2
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