Abstract
The functioning of NAD(P)H:FMN‑oxidoreductase (Red) from Vibrio fischeri under conditions of macromolecular crowding (MMC) simulated in vitro by adding biopolymers (starch and gelatin) was studied. The dissociation rate constants and the activation energies of dissociation of Red to the subunits were calculated, and the process of denaturation of Red was analyzed. It is shown that the functioning of Red both under conditions of MMC and in diluted solutions is the same. This result refutes the common belief that the native conformation of enzymes in vivo is stabilized due to MMC as compared to the in vitro conditions.
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Funding
The reported study was funded by Russian Foundation for Basic Research, Government of Krasnoyarsk Territory, Krasnoyarsk Regional Fund of Science as part of the research projects nos. 18-44‑243010 “Stabilization of coupled enzyme system of luminescent bacteria NADH:FMN-oxidoreductase + luciferase by placing into gels and viscous solutions of biopolymers in order to improve the reagent used for environmental contamination level assessment” and 18-44-242003 “Construction of an Enzymatic Reagent for Bioluminescent Analysis: Mechanisms to Increase the Sensitivity and Accuracy.”
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Translated by M. Batrukova
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Govorun, A.E., Esimbekova, E.N. & Kratasyuk, V.A. NAD(P)H:FMN‑Oxidoreductase Functioning Under Macromolecular Crowding: In Vitro Modeling. Dokl Biochem Biophys 486, 213–215 (2019). https://doi.org/10.1134/S160767291903013X
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DOI: https://doi.org/10.1134/S160767291903013X