Figure - available from: International Journal of Molecular Sciences (IJMS)
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In vitro enzyme assays were analyzed with Malon. (A) Schematic of the enzyme reactions being assessed. Fumarase converts fumarate to malate, and malate dehydrogenase (MDH) oxidizes malate to oxaloacetate by reducing NAD⁺ to NADH, or vice versa. (B) Malon fluorescence in enzyme reactions with fumarate and 0, 100 nM, or 1 μM fumarase, compared to direct detection of 15 mM malate. (C) Malon fluorescence in enzyme reactions with 0 or 1 μM MDH pre-equilibrated with malate upon NAD⁺ injection. (D) Same as part C, except pre-equilibrated with oxaloacetate upon NADH injection.
Source publication
Malate is a key intermediate in the citric acid cycle, an enzymatic cascade that is central to cellular energy metabolism and that has been applied to make biofuel cells. To enable real-time sensing of malate levels, we have engineered a genetically encoded, protein-based fluorescent biosensor called Malon specifically responsive to malate by perfo...
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