“4D MAPPING OF SPATIOFUNCTIONAL ENZYME DROPLETS IN LIVING CELLS”
Minjoung Kyoung, Assistant Professor of Chemistry and Biochemistry, University of Maryland
Abstract: The cytoplasmic, rate-determining enzymes in glucose metabolism are spatially organized into multienzyme assemblies in various sizes in human cells. They are proposed to shunt metabolic flux from glycolysis to anabolic biosynthetic pathways. However, how the formation of the enzyme assemblies and their sub-cellular locations in the cells are associated with their metabolic functions are largely unknown. In this work, we show that the enzyme assemblies in glucose metabolism are formed by liquid phase separation in a pathway specific manner. With 4D imaging using the home-built lattice light sheet microscope, we observe that the membraneless assemblies present liquid droplet properties. Moreover, we reveal that significant numbers of the enzyme assemblies are in proximity with mitochondria. We also monitor the reversible formation and spatial organization of the enzyme assembly upon the inhibition of mitochondrial metabolism. Our results shed light on how the enzyme assemblies are formed and positioned near cellular organelles to locally and efficiently orchestrate their metabolic functions. We envision that the presented “spatiofunctional” characteristics of the enzyme assemblies in glucose metabolism are an unprecedented starting point for mapping 4D functional metabolic network in live cells.
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