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Summary
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A novel stressor-actuated proximity labeling (SAPL) strategy uses host cell-released hydrogen peroxide (H2O2) to monitor cell-cell interactions. This method effectively records cellular stress levels, differentiating macrophage phenotypes and monitoring surface modification effects.

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Area of Science:

  • Cellular and Molecular Biology
  • Immunology
  • Biotechnology

Background:

  • Cell-cell interactions are crucial for determining cellular activation and function.
  • Host cells release hydrogen peroxide (H2O2) in response to stressors, indicating cellular stress.
  • Peroxidase enzymes can catalyze proximity labeling reactions when H2O2 is present.

Purpose of the Study:

  • To develop a novel stressor-actuated proximity labeling (SAPL) strategy.
  • To report on cell-cell interaction processes by recording cellular stress levels.
  • To utilize endogenous effector molecules for monitoring cellular interactions.

Main Methods:

  • Stressors were covalently modified with horseradish peroxidase (HRP).
  • Host cells' endogenous H2O2 release in response to stressors triggered HRP-based proximity labeling.
  • In situ imaging of labeling signals was used to compare stress levels, employing fungal mimics or live fungi as stressors.

Main Results:

  • SAPL effectively recorded stress levels during cell-cell interactions.
  • The strategy demonstrated enhanced sensitivity in differentiating interactions involving various macrophage phenotypes.
  • SAPL enabled real-time, in situ monitoring of surface modification impacts on cellular interactions.

Conclusions:

  • The SAPL strategy provides a new method for monitoring cell-cell interactions.
  • SAPL leverages endogenous H2O2 to report on cellular stress and interaction dynamics.
  • This approach offers a sensitive and versatile tool for studying cellular communication.