Biosensors-Based In Vivo Quantification of 2-Oxoglutarate in Cyanobacteria and Proteobacteria

2-oxoglutarate (α-ketoglutarate; 2-OG) is an intermediate of the Krebs cycle, and constitutes the carbon skeleton for nitrogen assimilation and the synthesis of a variety of compounds. In addition to being an important metabolite, 2-OG is a signaling molecule with a broad regulatory repertoire in a...

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Bibliographic Details
Published in:Artificial life 2018-10, Vol.8 (4), p.51
Main Authors: Chen, Hai-Lin, Latifi, Amel, Zhang, Cheng-Cai, Bernard, Christophe Sébastien
Format: Article
Language:English
Subjects:
2-oxoglutarate
Anabaena
Antibiotics
Bacteria
Binding sites
biosensor
Biosensors
Carbon
Communication
Cyanobacteria
E coli
Energy transfer
Escherichia coli
Fluorescence
Fluorescence resonance energy transfer
FRET
Gene expression
Ketoglutaric acid
Kinases
Krebs cycle
Life Sciences
Metabolism
Metabolites
Mutagenesis
Mutation
Nitrogen
Organisms
Plasmids
Proteins
RNA polymerase
Signal transduction
Technology transfer
Tricarboxylic acid cycle
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Summary:2-oxoglutarate (α-ketoglutarate; 2-OG) is an intermediate of the Krebs cycle, and constitutes the carbon skeleton for nitrogen assimilation and the synthesis of a variety of compounds. In addition to being an important metabolite, 2-OG is a signaling molecule with a broad regulatory repertoire in a variety of organisms, including plants, animals, and bacteria. Although challenging, measuring the levels and variations of metabolic signals in vivo is critical to better understand how cells control specific processes. To measure cellular 2-OG concentrations and dynamics, we designed a set of biosensors based on the fluorescence resonance energy transfer (FRET) technology that can be used in vivo in different organisms. For this purpose, we took advantage of the conformational changes of two cyanobacterial proteins induced by 2-OG binding. We show that these biosensors responded immediately and specifically to different 2-OG levels, and hence allowed to measure 2-OG variations in function of environmental modifications in the proteobacterium and in the cyanobacterium sp. PCC 7120. Our results pave the way to study 2-OG dynamics at the cellular level in uni- and multi-cellular organisms.
ISSN:2075-1729
1064-5462
2075-1729
1530-9185
DOI:10.3390/life8040051