Related Experiment Video

Updated: Jun 13, 2025

Fluorescence-based Monitoring of PAD4 Activity via a Pro-fluorescence Substrate Analog
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Fluorescence-based Monitoring of PAD4 Activity via a Pro-fluorescence Substrate Analog

Published on: November 5, 2014

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Evolutionary and molecular basis of ADP-ribosylation reversal by zinc-dependent macrodomains

Antonio Ariza1, Qiang Liu2, Nathan P Cowieson3

  • 1School of Biosciences, University of Sheffield, Sheffield, UK; Sir William Dunn School of Pathology, University of Oxford, Oxford, UK.

The Journal of Biological Chemistry
|September 13, 2024
PubMed

Insights

Zinc-containing macrodomains (Zn-Macros) are key in microbial defense against oxidative stress. This study reveals their catalytic mechanisms and substrate selectivity, offering potential for new antimicrobial therapies.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Structural Biology

Background:

  • ADP-ribosylation signaling regulates cellular responses to stress, including DNA damage and infection.
  • Pathogenic microbes utilize SirTM/zinc-containing macrodomain (Zn-Macro) enzyme pairs to manage oxidative stress responses.
  • Understanding Zn-Macro function is crucial for developing strategies against antimicrobial resistance.

Purpose of the Study:

  • To elucidate the functional properties of Zn-Macro enzymes involved in microbial defense.
  • To investigate the catalytic role of the zinc ion in ADP-ribosyl modification removal.
  • To determine structural determinants of substrate selectivity within the Zn-Macro family.

Main Methods:

  • Phylogenetic analysis to understand evolutionary relationships.
  • Biochemical assays using substrate mimetics and product analogs.
  • Structural biology approaches to determine enzyme-host interactions.

Main Results:

  • Characterization of the catalytic role of zinc ions in the removal of ADP-ribosyl modifications.
  • Identification of structural features governing substrate selectivity across different Zn-Macro branches.
  • Elucidation of the functional mechanisms of Zn-Macro enzymes in microbial stress response.

Conclusions:

  • Zn-Macro enzymes play a critical role in microbial defense mechanisms.
  • The zinc ion is essential for the catalytic activity of Zn-Macros in removing ADP-ribosylation.
  • Distinct structural features of Zn-Macros contribute to their substrate specificity, presenting therapeutic targets.
Keywords:
MetalloenzymePost-translational modification (PTM)X-ray crystallographylipoylationsmall-angle X-ray scattering (SAXS)

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