Sharrock, A.V.; Mumm, J.S.; Bagdžiūnas, G.; Čėnas, 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.2023, 24, 6633.
Sharrock, A.V.; Mumm, J.S.; Bagdžiūnas, G.; Čėnas, 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. 2023, 24, 6633.
Sharrock, A.V.; Mumm, J.S.; Bagdžiūnas, G.; Čėnas, 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.2023, 24, 6633.
Sharrock, A.V.; Mumm, J.S.; Bagdžiūnas, G.; Čėnas, 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. 2023, 24, 6633.
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 Vibrio vulnificus. 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.
Keywords
Type I nitroreductase; NfsB; Crystal structure; Prodrug; Metronidazole; Targeted cellular ablation; Tinidazole; CB1954; Dinitrotoluene
Subject
Biology and Life Sciences, Biochemistry and Molecular Biology
Copyright:
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
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