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Volume 169, Issue 1

Assay development and inhibition of the -DprE2 essential reductase from Open Access

This article is part of the Diversity in Microbiology collection

Abstract

DprE2 is an essential enzyme in the synthesis of decaprenylphosphoryl-β--arabinofuranose (DPA) and subsequently arabinogalactan, and is a significant new drug target for . Two compounds from the GSK-177 box set, GSK301A and GSK032A, were identified through -DprE2-target overexpression studies. The -DprE1-DprE2 complex was co-purified and a new DprE2 assay developed, based on the oxidation of the reduced nicotinamide adenine dinucleotide cofactor of DprE2 (NADH/NADPH). The -DprE1-DprE2 complex showed interesting kinetics in both the DprE1 resazurin-based assay, where -DprE2 was found to enhance -DprE1 activity and reduce substrate inhibition; and also in the DprE2 assay, which similarly exhibited substrate inhibition and a difference in kinetics of the two potential cofactors, NADH and NADPH. Although, no inhibition was observed in the DprE2 assay by the two GSK set compounds, spontaneous mutant generation indicated a possible explanation in the form of a pro-drug activation pathway, involving and .

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Keyword(s): decaprenylphosphoryl-D-arabinose , enzyme kinetics , Mt-DprE1 , Mt-DprE2 , mycobacterial cell wall , Mycobacterium tuberculosis and substrate inhibition
Funding
This study was supported by the:
  • Medical Research Foundation (Award MR/R001154/1)
    • Principal Award Recipient: GurdyalS. Besra
  • Medical Research Foundation (Award MR/S000542/1)
    • Principal Award Recipient: GurdyalS. Besra
© 2023 The Authors
  • This is an open-access article distributed under the terms of the Creative Commons Attribution License. This article was made open access via a Publish and Read agreement between the Microbiology Society and the corresponding author’s institution.
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2023-01-23
2026-02-19

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/content/journal/micro/10.1099/mic.0.001288
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Assay development and inhibition of the Mt-DprE2 essential reductase from Mycobacterium tuberculosis
Microbiology 169, 001288 (2023); https://doi.org/10.1099/mic.0.001288
/content/journal/micro/10.1099/mic.0.001288
/content/journal/micro/10.1099/mic.0.001288
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