Enzymatic analysis of Glucose-1-Phosphate Adenylyltransferase (GlgC) reveals tight metabolite-level regulation of glycogen synthesis in cyanobacteria

Abstract The enzyme Glucose-1-phosphate adenylyltransferase (GlgC, EC:2.7.7.27) catalyses the first step in glycogen synthesis by converting glucose-1-phosphate into ADP-Glucose, which is added in turn to a growing glycogen chain by glycogen synthases. Thus far, the in vitro study of GlgC were mainly performed using colorimetric or radiolabel-based phosphate release assays, limiting the option for analysing this reaction. With this work, we present a novel in vitro continuous assay coupling the subsequent glycogen synthase reaction to the GlgC reaction, thus simulating the process of glycogen synthesis in vivo. Using this assay, we revisited GlgC catalytic parameters and screened for metabolites that affect GlgC activity in Synechocystis sp. PCC 6803. We also describe in further detail the antagonistic interplay between the GlgC activator, 3-PGA and the inhibitor, inorganic phosphate, revealing the intricate mechanism by which glycogen formation responds to fluctuations in carbon and energy supply in cyanobacteria.

Here you will find data files for the results presented in this work, in Excel (.xlsx) and Graphpad (.pzf) format.

DOI: 10.15490/fairdomhub.1.study.1296.1

Zenodo URL: None

Created at: 5th Jul 2024 at 19:40

Contents

GlgC coupled assay

No description specified

GlgC Experimental Data

Selected data sets from the main figures presented in this work.

  • GlgC Paper Data (selected).xlsx

GlgC Figure Data

All data sets reflected by the figures in this work. Requires Graphpad Prism 7 (or later) to access. A trial version can be downloaded from the Graphpad prism website ( https://www.graphpad.com/demos/ )

  • GlgC Paper Data and figures (all).pzf
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Citation
Lee, K. (2024). Enzymatic analysis of Glucose-1-Phosphate Adenylyltransferase (GlgC) reveals tight metabolite-level regulation of glycogen synthesis in cyanobacteria. FAIRDOMHub. https://doi.org/10.15490/FAIRDOMHUB.1.STUDY.1296.1
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