7UWT

Structure of Oxygen-Insensitive NAD(P)H-dependent Nitroreductase NfsB_Vv F70A/F108Y (NTR 2.0) in complex with FMN at 1.85 Angstroms resolution

Summary for 7UWT

• Entry DOI: 10.2210/pdb7uwt/pdb
• Descriptor: Dihydropteridine reductase, FLAVIN MONONUCLEOTIDE, ACETATE ION, ... (5 entities in total)
• Functional Keywords: nitroreductase, oxidoreductase, flavoenzyme, metronidazole reductase
• Biological source: Vibrio vulnificus
• Total number of polymer chains: 2
• Total formula weight: 51317.88

Authors

Sharrock, A.V.,Arcus, V.,Mumm, J.S.,Ackerley, D.F. (deposition date: 2022-05-03, release date: 2022-05-18, Last modification date: 2023-10-25)

Primary citation

Sharrock, A.V.,Mumm, J.S.,Bagdziunas, G.,Cenas, N.,Arcus, V.L.,Ackerley, D.F.
The Crystal Structure of Engineered Nitroreductase NTR 2.0 and Impact of F70A and F108Y Substitutions on Substrate Specificity.
Int J Mol Sci, 24:-, 2023

PubMed Abstract: Bacterial nitroreductase enzymes that convert prodrugs to cytotoxins are valuable tools for creating transgenic targeted ablation models to study cellular function and cell-specific regeneration paradigms. We recently engineered a nitroreductase ("NTR 2.0") for substantially enhanced reduction of the prodrug metronidazole, which permits faster cell ablation kinetics, cleaner interrogations of cell function, ablation of previously recalcitrant cell types, and extended ablation paradigms useful for modelling chronic diseases. To provide insight into the enhanced enzymatic mechanism of NTR 2.0, we have solved the X-ray crystal structure at 1.85 Angstroms resolution and compared it to the parental enzyme, NfsB from . We additionally present a survey of reductive activity with eight alternative nitroaromatic substrates, to provide access to alternative ablation prodrugs, and explore applications such as remediation of dinitrotoluene pollutants. The predicted binding modes of four key substrates were investigated using molecular modelling.

PubMed: 37047605
DOI: 10.3390/ijms24076633

Experimental method

X-RAY DIFFRACTION (1.85 Å)