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DNA probes are fragments of DNA labeled with a reporter tag to enable their detection or purification. The resulting labeled DNA probes can then hybridize to target nucleic acid sequences through complementary base-pairing, and may be used to recover or identify these regions.
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In situ hybridization (ISH) is a technique used to detect and localize specific DNA or RNA molecules in cells, tissue, or tissue sections using a labeled probe. The technique was first used in 1969 for the investigation of nucleic acids. It is currently an essential tool in scientific research and clinical settings, especially for diagnostic purposes.
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  1. Home
  3. Bespoke Activity-based Probes Reveal That The Pseudomonas Aeruginosa Endoglycosidase, Pslg, Is An Endo-β-glucanase.
  1. Home
  3. Bespoke Activity-based Probes Reveal That The Pseudomonas Aeruginosa Endoglycosidase, Pslg, Is An Endo-β-glucanase.

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Bespoke Activity-Based Probes Reveal that the Pseudomonas aeruginosa Endoglycosidase, PslG, Is an Endo-β-glucanase.

Gijs Ruijgrok1, Wendy A Offen2, Isabelle B Pickles2

  • 1Leiden Institute of Chemistry, Leiden University, 2300 RA Leiden, The Netherlands.

Journal of the American Chemical Society
|February 25, 2025

View abstract on PubMed

Summary
This summary is machine-generated.

Pseudomonas aeruginosa forms biofilms using Psl, a sugar matrix. Researchers discovered the enzyme PslG is an endo-β-glucosidase, not a mannosidase, enabling new inhibitor designs to combat antibiotic resistance.

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Area of Science:

  • Microbiology
  • Biochemistry
  • Structural Biology

Background:

  • Pseudomonas aeruginosa forms biofilms crucial for infection persistence.
  • Biofilm matrix polysaccharides, like Psl, provide protection against antibiotics and host immunity.
  • The Psl biosynthesis pathway includes the enzyme PslG, whose function was previously misidentified.

Purpose of the Study:

  • To elucidate the enzymatic activity of PslG involved in Psl matrix biosynthesis.
  • To characterize PslG as a potential target for inhibiting biofilm formation.
  • To develop novel inhibitors for PslG to combat antibiotic resistance in P. aeruginosa.

Main Methods:

  • Activity-based protein profiling to identify enzyme function.
  • Structural studies of enzyme-inhibitor complexes.
  • Defined substrate processing assays.
  • Design and synthesis of PslG inhibitors.

    • Main Results:

    • PslG was identified as a retaining endo-β-glucosidase, not an endo-β-mannosidase.
    • Structural insights enabled the design of competitive and covalent PslG inhibitors.
    • Inhibitors based on deoxynojirimycin and cyclophellitol moieties were successfully designed.

    Conclusions:

    • PslG's true endo-β-glucosidase activity is critical for Psl biosynthesis.
    • This discovery provides tools to study Psl function and biofilm development.
    • The developed inhibitors offer a strategy to target P. aeruginosa biofilms and enhance antibiotic efficacy.