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Updated: Jun 13, 2026

Isolation of Translating Ribosomes Containing Peptidyl-tRNAs for Functional and Structural Analyses
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Time-Dependent Inhibition of Leucyl-tRNA-Synthetase (LeuRS): Insight into Target Vulnerability.

Mingqian Wang1, YongLe He1, Siobhan A Cohen1

  • 1Center for Advanced Study of Drug Action, and Department of Chemistry, Stony Brook University, Stony Brook, New York 11794-3400, United States.

ACS Infectious Diseases
|April 2, 2025
PubMed
Summary

Benzoxaboroles inhibit bacterial protein synthesis by targeting leucyl-tRNA synthetase. Combining these with tobramycin enhances the postantibiotic effect (PAE) by reducing enzyme synthesis, offering new dosing strategies.

Keywords:
epetraboroleleucyl-tRNA-synthetasepSILACpostantibiotic effectresidence timetarget vulnerability

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Area of Science:

  • Microbiology
  • Pharmacology
  • Biochemistry

Background:

  • The postantibiotic effect (PAE) is crucial for optimizing antimicrobial dosing regimens.
  • Understanding drug-target interactions and protein turnover is key to elucidating PAE mechanisms.

Purpose of the Study:

  • To investigate the postantibiotic effect (PAE) of benzoxaboroles targeting *Escherichia coli* leucyl-tRNA synthetase (ecLeuRS).
  • To assess the impact of tobramycin on benzoxaborole-induced PAE and ecLeuRS synthesis.

Main Methods:

  • Time-dependent inhibition assays were used to study ecLeuRS inhibition by benzoxaboroles.
  • Postantibiotic effect (PAE) was measured in *E. coli* with and without sub-inhibitory concentrations of tobramycin.
  • pSILAC was employed to quantify changes in ecLeuRS synthesis rates.

Main Results:

  • Benzoxaboroles, including epetraborole, showed slow-binding inhibition of ecLeuRS, significantly reducing IC50 values.
  • Short PAEs (<1 h) were observed for benzoxaboroles alone, but increased to 1.70-3 h when combined with tobramycin.
  • Sub-inhibitory tobramycin reduced ecLeuRS synthesis by 1.6-fold, indicating a role for protein turnover in target vulnerability.

Conclusions:

  • Benzoxaboroles effectively inhibit ecLeuRS, with enhanced binding affinity observed.
  • The combination of benzoxaboroles and tobramycin potentiates the PAE, suggesting synergistic effects.
  • Protein turnover significantly influences target vulnerability and the overall postantibiotic effect, informing future antimicrobial drug development.