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Multiple Glycogen-binding Sites in Eukaryotic Glycogen Synthase are Required for High Catalytic Efficiency Towards Glycogen

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71a08xRcMcL.jpgGlycogen synthase is a rate-limiting enzyme in the biosynthesis of glycogen and has an important role in glucose homeostasis. The three-dimensional constructions of yeast glycogen synthase (Gsy2p) complexed with maltooctaose recognized 4 conserved maltodextrin-binding sites distributed across the surface of the enzyme. Site-1 is positioned on the N-terminal domain, site-2 and site-three are present on the C-terminal area, and site-four is situated in an interdomain cleft adjacent to the active site. Mutation of those surface sites decreased glycogen binding and catalytic effectivity toward glycogen. 40- and 70-fold, improve energy levels respectively. 1-gsy2) remodeled with the site-1, site-2, combined site-1/site-2, or site-four mutant type of Gsy2p was decreased by up to 40-fold. In contrast to the glycogen results, the ability to utilize maltooctaose as an in vitro substrate was unaffected in the site-2 mutant, moderately affected in the positioning-1 mutant, and nearly utterly abolished in the positioning-4 mutant. These knowledge show that the power to make the most of maltooctaose as a substrate could be impartial of the power to utilize glycogen. Our knowledge support the speculation that site-1 and site-2 provide a 'toehold mechanism,' preserving glycogen synthase tightly related to the glycogen particle, whereas site-4 is extra closely related to positioning of the nonreducing end during catalysis.

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