Structural Insight into Catalysis by the Flavin-Dependent NADH Oxidase (Pden_5119) of Paracoccus denitrificans

Investor logo
Investor logo

Warning

This publication doesn't include Institute of Computer Science. It includes Faculty of Science. Official publication website can be found on muni.cz.
Authors

KRYL Martin SEDLÁČEK Vojtěch KUČERA Igor

Year of publication 2023
Type Article in Periodical
Magazine / Source International Journal of Molecular Sciences
MU Faculty or unit

Faculty of Science

Citation
Web https://www.mdpi.com/1422-0067/24/4/3732
Doi http://dx.doi.org/10.3390/ijms24043732
Keywords FMN; NADH; dioxygen reduction; Paracoccus denitrificans
Description The Pden_5119 protein oxidizes NADH with oxygen under mediation by the bound flavin mononucleotide (FMN) and may be involved in the maintenance of the cellular redox pool. In biochemical characterization, the curve of the pH-rate dependence was bell-shaped with pK(a1) = 6.6 and pK(a2) = 9.2 at 2 mu M FMN while it contained only a descending limb pK(a) of 9.7 at 50 mu M FMN. The enzyme was found to undergo inactivation by reagents reactive with histidine, lysine, tyrosine, and arginine. In the first three cases, FMN exerted a protective effect against the inactivation. X-ray structural analysis coupled with site-directed mutagenesis identified three amino acid residues important to the catalysis. Structural and kinetic data suggest that His-117 plays a role in the binding and positioning of the isoalloxazine ring of FMN, Lys-82 fixes the nicotinamide ring of NADH to support the proS-hydride transfer, and Arg-116 with its positive charge promotes the reaction between dioxygen and reduced flavin.
Related projects:

You are running an old browser version. We recommend updating your browser to its latest version.

More info