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Updated: Jun 26, 2025
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Genome-wide Protein-protein Interaction Screening by Protein-fragment Complementation Assay PCA in Living Cells
Published on: March 3, 2015
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A Semiquantitative Protein-Fragment Complementation Assay to Study Protein-Protein Interactions of the Polymerase
Joanna Brunel1, Érica Urzua1, Denis Gerlier1
1Centre International de Recherche en Infectiologie (CIRI), Inserm U1111, CNRS UMR5308, Université Claude Bernard Lyon1, ENS Lyon, Lyon, France.
Methods in Molecular Biology (Clifton, N.J.)
|May 14, 2024
Summary
Protein-fragment complementation assays (PCAs) reveal measles virus polymerase interactions. The split-luciferase assay effectively maps viral protein partnerships in cells.
Area of Science:
- Molecular Biology
- Virology
- Biochemistry
Background:
- Protein-fragment complementation assays (PCAs) are vital for studying protein interactions within cells.
- PCAs are particularly useful for analyzing unstable proteins and weak interactions.
- The Gaussia princeps luciferase (split-luc) PCA offers high sensitivity for interaction mapping and affinity measurement.
Purpose of the Study:
- To describe a split-luciferase protocol for mapping protein-protein interactions.
- To investigate the viral interactome of the measles virus polymerase complex.
Main Methods:
- Utilized a split-luciferase protein-fragment complementation assay (split-luc PCA).
- Applied the assay to map interactions within the measles virus polymerase complex.
- Focused on interactions that are difficult to study using traditional biochemical methods.
Main Results:
- Successfully mapped protein-protein interactions of the measles virus polymerase complex.
- Demonstrated the utility of the split-luc PCA for studying viral protein partnerships.
- Provided a detailed protocol for future interactome studies.
Conclusions:
- The split-luc PCA is a powerful and sensitive method for mapping viral interactomes.
- This approach enables the study of challenging protein interactions in a native cellular environment.
- The described protocol facilitates the investigation of measles virus polymerase complex interactions.
