Optimization of a Bioluminescence Resonance Energy Transfer-Based Assay for Screening of Trypanosoma cruzi Protein/Protein Interaction Inhibitors

Mol Biotechnol. 2018 May;60(5):369-379. doi: 10.1007/s12033-018-0078-3.

Abstract

Chagas disease, a parasitic disease caused by Trypanosoma cruzi, is a major public health burden in poor rural populations of Central and South America and a serious emerging threat outside the endemic region, since the number of infections in non-endemic countries continues to rise. In order to develop more efficient anti-trypanosomal treatments to replace the outdated therapies, new molecular targets need to be explored and new drugs discovered. Trypanosoma cruzi has distinctive structural and functional characteristics with respect to the human host. These exclusive features could emerge as interesting drug targets. In this work, essential and differential protein-protein interactions for the parasite, including the ribosomal P proteins and proteins involved in mRNA processing, were evaluated in a bioluminescence resonance energy transfer-based assay as a starting point for drug screening. Suitable conditions to consider using this simple and robust methodology to screening compounds and natural extracts able to inhibit protein-protein interactions were set in living cells and lysates.

Keywords: BRET; Drug screening; Protein–protein interactions; Trypanosoma cruzi.

MeSH terms

  • Central America
  • Drug Evaluation, Preclinical
  • Fluorescence Resonance Energy Transfer / methods*
  • HEK293 Cells
  • Humans
  • Protein Binding / drug effects
  • Protein Interaction Maps
  • Protozoan Proteins / chemistry
  • Protozoan Proteins / metabolism*
  • Small Molecule Libraries / pharmacology
  • South America
  • Trypanocidal Agents / pharmacology*
  • Trypanosoma cruzi / drug effects*
  • Trypanosoma cruzi / metabolism

Substances

  • Protozoan Proteins
  • Small Molecule Libraries
  • Trypanocidal Agents