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Visualization of Bacterial Resistance using Fluorescent Antibiotic Probes
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An Activity-Based Probe for Studying Crosslinking in Live Bacteria.

Samir Gautam1,2, Taehan Kim2, Takuji Shoda2,3

  • 1Department of Cell Biology, Yale School of Medicine, 333 Cedar Street, New Haven, CT 06520 (USA).

Angewandte Chemie (International Ed. in English)
|July 25, 2015
PubMed
Summary
This summary is machine-generated.

Scientists developed a new tool to visualize bacterial cell wall crosslinking in real-time. This fluorescent probe specifically labels penicillin-binding protein 4 (PBP4) activity in Staphylococcus aureus, revealing its location during cell division.

Keywords:
bacteriabiosensorscrosslinkingfluorescent probespeptidoglycans

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Area of Science:

  • Microbiology
  • Biochemistry
  • Cell Biology

Background:

  • Penicillin-binding proteins (PBPs) are crucial for bacterial cell wall synthesis and are key antibiotic targets.
  • Understanding the spatiotemporal dynamics of peptidoglycan crosslinking is vital but hindered by a lack of experimental tools.
  • Current methods do not allow for real-time visualization of PBP activity in live bacteria.

Purpose of the Study:

  • To develop and validate a novel activity-based probe for visualizing PBP-mediated peptidoglycan crosslinking in live bacteria.
  • To investigate the spatiotemporal localization and activity of PBPs in Staphylococcus aureus.
  • To identify specific PBPs targeted by the developed probe.

Main Methods:

  • Development of fluorescent stem peptide mimics (FSPMs) as activity-based probes.
  • Application of FSPMs for pulse-labeling and visualization of PBP activity in live Staphylococcus aureus.
  • Quantitation of FSPM incorporation into the bacterial cell wall.
  • Localization studies of PBP activity using FSPM labeling.

Main Results:

  • Fluorescent stem peptide mimics (FSPMs) were successfully synthesized and demonstrated to be covalently incorporated into the cell wall of Staphylococcus aureus.
  • FSPMs selectively target and are recognized by a single PBP in S. aureus, identified as PBP4.
  • FSPM pulse-labeling enabled the visualization and relative quantitation of PBP4 activity in vivo.
  • PBP4 activity was localized to the septum during cell division in live S. aureus cells.
  • PBP4 recruitment to the septum was shown to be dependent on wall teichoic acid.

Conclusions:

  • A novel activity-based probe (FSPM) has been developed, enabling real-time visualization and study of PBP activity in live bacteria.
  • The probe selectively labels PBP4 in Staphylococcus aureus, providing insights into its specific role and localization.
  • PBP4 activity is dynamically regulated and localized to the septum, dependent on wall teichoic acid, during bacterial growth and division.