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

Publication ,  Journal Article
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 in: ACS Infect Dis
April 12, 2019

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.

Duke Scholars

Published In

ACS Infect Dis

DOI

EISSN

2373-8227

Publication Date

April 12, 2019

Volume

5

Issue

4

Start / End Page

641 / 651

Location

United States

Related Subject Headings

  • Structure-Activity Relationship
  • Rosaniline Dyes
  • Pyrophosphatases
  • Piperazine
  • Phosphates
  • Glycolipids
  • Escherichia coli
  • Enzyme Inhibitors
  • Enzyme Assays
  • Biocatalysis
 

Citation

APA
Chicago
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MLA
NLM
Lee, M., Zhao, J., Kwak, S.-H., Cho, J., Gillespie, R. A., Kwon, D.-Y., … Hong, J. (2019). Structure-Activity Relationship of Sulfonyl Piperazine LpxH Inhibitors Analyzed by an LpxE-Coupled Malachite Green Assay. ACS Infect Dis, 5(4), 641–651. https://doi.org/10.1021/acsinfecdis.8b00364
Lee, Minhee, Jinshi Zhao, Seung-Hwa Kwak, Jae Cho, Myungju Lee, Robert A. Gillespie, Do-Yeon Kwon, et al. “Structure-Activity Relationship of Sulfonyl Piperazine LpxH Inhibitors Analyzed by an LpxE-Coupled Malachite Green Assay.ACS Infect Dis 5, no. 4 (April 12, 2019): 641–51. https://doi.org/10.1021/acsinfecdis.8b00364.
Lee M, Zhao J, Kwak S-H, Cho J, Gillespie RA, Kwon D-Y, et al. Structure-Activity Relationship of Sulfonyl Piperazine LpxH Inhibitors Analyzed by an LpxE-Coupled Malachite Green Assay. ACS Infect Dis. 2019 Apr 12;5(4):641–51.
Lee, Minhee, et al. “Structure-Activity Relationship of Sulfonyl Piperazine LpxH Inhibitors Analyzed by an LpxE-Coupled Malachite Green Assay.ACS Infect Dis, vol. 5, no. 4, Apr. 2019, pp. 641–51. Pubmed, doi:10.1021/acsinfecdis.8b00364.
Lee M, Zhao J, Kwak S-H, Cho J, Gillespie RA, Kwon D-Y, Lee H, Park H-J, Wu Q, Zhou P, Hong J. Structure-Activity Relationship of Sulfonyl Piperazine LpxH Inhibitors Analyzed by an LpxE-Coupled Malachite Green Assay. ACS Infect Dis. 2019 Apr 12;5(4):641–651.

Published In

ACS Infect Dis

DOI

EISSN

2373-8227

Publication Date

April 12, 2019

Volume

5

Issue

4

Start / End Page

641 / 651

Location

United States

Related Subject Headings

  • Structure-Activity Relationship
  • Rosaniline Dyes
  • Pyrophosphatases
  • Piperazine
  • Phosphates
  • Glycolipids
  • Escherichia coli
  • Enzyme Inhibitors
  • Enzyme Assays
  • Biocatalysis