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Updated: Sep 16, 2025

Characterization of Neuronal Lysosome Interactome with Proximity Labeling Proteomics
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Exploring Bacterial Surface Proteome Dynamics During Infection Using Proximity Labeling.

Oceane Dubois1, Abdelrahim Zoued2

  • 1CIRI, Centre International de Recherche en Infectiologie, CNRS UMR5308, Inserm U1111, Université Claude Bernard Lyon 1, ENS de Lyon, Lyon, France.

Methods in Molecular Biology (Clifton, N.J.)
|July 10, 2025
PubMed
Summary
This summary is machine-generated.

This study introduces proximity-dependent protein biotinylation to map bacterial surface proteins and host-binding proteins during infection. This method enhances understanding of host-pathogen interactions and identifies potential therapeutic targets.

Keywords:
APEX2AutotransporterBacterial surface labelingHost-derived bacterial-binding protein (HBBP)Host–bacteria interactionsType V Secretion System (T5SS)

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Area of Science:

  • Microbiology
  • Molecular Biology
  • Infectious Diseases

Background:

  • Studying bacterial surface proteins during infection is crucial for understanding host-pathogen interactions.
  • Existing methods may have limitations in precisely characterizing the bacterial surface proteome in situ.

Purpose of the Study:

  • To develop and validate a proximity-dependent protein biotinylation method for studying bacterial surface proteomes.
  • To identify bacterial surface proteins and host-derived bacterial-binding proteins (HBBPs) during infection.

Main Methods:

  • Engineering bacterial autotransporters to display APEX2 enzyme on the bacterial surface.
  • Implementing a workflow involving bacterial culturing, proximity-dependent biotinylation, and immunofluorescence labeling.
  • Utilizing protein analysis techniques to identify labeled proteins.

Main Results:

  • Successful labeling and characterization of bacterial surface proteins.
  • Identification of host-derived bacterial-binding proteins (HBBPs) interacting with the pathogen.
  • Demonstration of a comprehensive workflow for host-pathogen interaction studies.

Conclusions:

  • Surface-exposed proximity labeling provides a powerful tool for dissecting bacterial pathogenesis.
  • This approach can reveal novel virulence factors and therapeutic targets.
  • The method is adaptable to various bacterial species and infection models.