Structure-Activity Relationship of Sulfonyl Piperazine LpxH Inhibitors Analyzed by an LpxE-Coupled Malachite Green Assay.

Journal Article (Journal Article)

The UDP-2,3-diacylglucosamine pyrophosphatase LpxH in the Raetz pathway of lipid A biosynthesis is an essential enzyme in the vast majority of Gram-negative pathogens and an excellent novel antibiotic target. The 32P-radioautographic thin-layer chromatography assay has been widely used for analysis of LpxH activity, but it is inconvenient for evaluation of a large number of LpxH inhibitors over an extended time period. Here, we report a coupled, nonradioactive LpxH assay that utilizes the recently discovered Aquifex aeolicus lipid A 1-phosphatase LpxE for quantitative removal of the 1-phosphate from lipid X, the product of the LpxH catalysis; the released inorganic phosphate is subsequently quantified by the colorimetric malachite green assay, allowing the monitoring of the LpxH catalysis. Using such a coupled enzymatic assay, we report the biochemical characterization of a series of sulfonyl piperazine LpxH inhibitors. Our analysis establishes a preliminary structure-activity relationship for this class of compounds and reveals a pharmacophore of two aromatic rings, two hydrophobic groups, and one hydrogen-bond acceptor. We expect that our findings will facilitate the development of more effective LpxH inhibitors as potential antibacterial agents.

Full Text

Duke Authors

Cited Authors

  • Lee, M; Zhao, J; Kwak, S-H; Cho, J; Lee, M; Gillespie, RA; Kwon, D-Y; Lee, H; Park, H-J; Wu, Q; Zhou, P; Hong, J

Published Date

  • April 12, 2019

Published In

Volume / Issue

  • 5 / 4

Start / End Page

  • 641 - 651

PubMed ID

  • 30721024

Pubmed Central ID

  • PMC6730544

Electronic International Standard Serial Number (EISSN)

  • 2373-8227

Digital Object Identifier (DOI)

  • 10.1021/acsinfecdis.8b00364


  • eng

Conference Location

  • United States