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Inhibition of lipid A biosynthesis as the primary mechanism of CHIR-090 antibiotic activity in Escherichia coli.

Publication ,  Journal Article
Barb, AW; McClerren, AL; Snehelatha, K; Reynolds, CM; Zhou, P; Raetz, CRH
Published in: Biochemistry
March 27, 2007

The deacetylation of UDP-3-O-[(R)-3-hydroxymyristoyl]-N-acetylglucosamine (UDP-3-O-acyl-GlcNAc) by LpxC is the committed reaction of lipid A biosynthesis. CHIR-090, a novel N-aroyl-l-threonine hydroxamic acid, is a potent, slow, tight-binding inhibitor of the LpxC deacetylase from the hyperthermophile Aquifex aeolicus, and it has excellent antibiotic activity against Pseudomonas aeruginosa and Escherichia coli, as judged by disk diffusion assays. We now report that CHIR-090 is also a two-step slow, tight-binding inhibitor of E. coli LpxC with Ki = 4.0 nM, Ki* = 0.5 nM, k5 = 1.9 min-1, and k6 = 0.18 min-1. CHIR-090 at low nanomolar levels inhibits LpxC orthologues from diverse Gram-negative pathogens, including P. aeruginosa, Neisseria meningitidis, and Helicobacter pylori. In contrast, CHIR-090 is a relatively weak competitive and conventional inhibitor (lacking slow, tight-binding kinetics) of LpxC from Rhizobium leguminosarum (Ki = 340 nM), a Gram-negative plant endosymbiont that is resistant to this compound. The KM (4.8 microM) and the kcat (1.7 s-1) of R. leguminosarum LpxC with UDP-3-O-[(R)-3-hydroxymyristoyl]-N-acetylglucosamine as the substrate are similar to values reported for E. coli LpxC. R. leguminosarum LpxC therefore provides a useful control for validating LpxC as the primary target of CHIR-090 in vivo. An E. coli construct in which the chromosomal lpxC gene is replaced by R. leguminosarum lpxC is resistant to CHIR-090 up to 100 microg/mL, or 400 times above the minimal inhibitory concentration for wild-type E. coli. Given its relatively broad spectrum and potency against diverse Gram-negative pathogens, CHIR-090 is an excellent lead for the further development of new antibiotics targeting the lipid A pathway.

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Published In

Biochemistry

DOI

ISSN

0006-2960

Publication Date

March 27, 2007

Volume

46

Issue

12

Start / End Page

3793 / 3802

Location

United States

Related Subject Headings

  • Lipid A
  • Kinetics
  • Hydroxamic Acids
  • Gram-Negative Bacterial Infections
  • Gram-Negative Bacteria
  • Escherichia coli
  • Biochemistry & Molecular Biology
  • Anti-Bacterial Agents
  • Amidohydrolases
  • 3404 Medicinal and biomolecular chemistry
 

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Barb, A. W., McClerren, A. L., Snehelatha, K., Reynolds, C. M., Zhou, P., & Raetz, C. R. H. (2007). Inhibition of lipid A biosynthesis as the primary mechanism of CHIR-090 antibiotic activity in Escherichia coli. Biochemistry, 46(12), 3793–3802. https://doi.org/10.1021/bi6025165
Barb, Adam W., Amanda L. McClerren, Karnem Snehelatha, C Michael Reynolds, Pei Zhou, and Christian R. H. Raetz. “Inhibition of lipid A biosynthesis as the primary mechanism of CHIR-090 antibiotic activity in Escherichia coli.Biochemistry 46, no. 12 (March 27, 2007): 3793–3802. https://doi.org/10.1021/bi6025165.
Barb AW, McClerren AL, Snehelatha K, Reynolds CM, Zhou P, Raetz CRH. Inhibition of lipid A biosynthesis as the primary mechanism of CHIR-090 antibiotic activity in Escherichia coli. Biochemistry. 2007 Mar 27;46(12):3793–802.
Barb, Adam W., et al. “Inhibition of lipid A biosynthesis as the primary mechanism of CHIR-090 antibiotic activity in Escherichia coli.Biochemistry, vol. 46, no. 12, Mar. 2007, pp. 3793–802. Pubmed, doi:10.1021/bi6025165.
Barb AW, McClerren AL, Snehelatha K, Reynolds CM, Zhou P, Raetz CRH. Inhibition of lipid A biosynthesis as the primary mechanism of CHIR-090 antibiotic activity in Escherichia coli. Biochemistry. 2007 Mar 27;46(12):3793–3802.
Journal cover image

Published In

Biochemistry

DOI

ISSN

0006-2960

Publication Date

March 27, 2007

Volume

46

Issue

12

Start / End Page

3793 / 3802

Location

United States

Related Subject Headings

  • Lipid A
  • Kinetics
  • Hydroxamic Acids
  • Gram-Negative Bacterial Infections
  • Gram-Negative Bacteria
  • Escherichia coli
  • Biochemistry & Molecular Biology
  • Anti-Bacterial Agents
  • Amidohydrolases
  • 3404 Medicinal and biomolecular chemistry