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Many proteins form complexes to carry out their functions, making protein-protein interactions (PPIs) essential for an organism's survival. Most PPIs are stabilized by numerous weak noncovalent chemical forces. The physical shape of the interfaces determines the way two proteins interact. Many globular proteins have closely-matching shapes on their surfaces, which form a large number of weak bonds. Additionally, many PPIs occur between two helices or between a surface cleft and a...
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An organism can have thousands of different proteins, and these proteins must cooperate to ensure the health of an organism. Proteins bind to other proteins and form complexes to carry out their functions. Many proteins interact with multiple other proteins creating a complex network of protein interactions.
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Updated: Sep 4, 2025

Investigating Protein-protein Interactions in Live Cells Using Bioluminescence Resonance Energy Transfer
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Screening of Protein-Protein Interaction Modulators Using BRET-Based Technology.

Lucia R Fernández1,2, Jesica Mild3,4, Martin M Edreira5,6,7

  • 1Universidad de Buenos Aires, Facultad de Ciencias Exactas y Naturales, Departamento de Química Orgánica, Buenos Aires, Argentina.

Methods in Molecular Biology (Clifton, N.J.)
|July 14, 2022
PubMed
Summary

We developed a bioluminescence resonance energy transfer (BRET) assay to screen for compounds that affect essential protein-protein interactions (PPIs) crucial for Trypanosoma cruzi survival. This method can analyze complex mixtures like natural extracts or chemical compounds.

Keywords:
Bioluminescence resonance energy transfer (BRET)Drug screeningInhibitorsModulatorsProtein–protein interactionTrypanosoma cruzi

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Area of Science:

  • Molecular Biology
  • Biochemistry
  • Parasitology

Background:

  • Protein-protein interactions (PPIs) are fundamental to cellular functions.
  • Bioluminescence resonance energy transfer (BRET) is a powerful technique for studying PPIs.
  • Trypanosoma cruzi survival depends on essential PPIs.

Purpose of the Study:

  • To establish a BRET-based assay for screening modulators of Trypanosoma cruzi PPIs.
  • To enable the identification of novel therapeutic targets for Chagas disease.

Main Methods:

  • Development of a BRET assay for PPI screening.
  • Application of BRET in living cells.
  • Application of BRET using cell lysates to analyze complex mixtures.

Main Results:

  • The BRET assay successfully screened for modulators of essential PPIs.
  • The assay is adaptable for both purified compounds and complex natural extracts.
  • Two distinct BRET approaches (living cells vs. lysates) were validated.

Conclusions:

  • The developed BRET assay is a versatile tool for identifying compounds that interfere with critical PPIs in Trypanosoma cruzi.
  • This assay facilitates drug discovery efforts against Chagas disease by enabling the screening of diverse chemical and natural product libraries.