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Related Experiment Video

Updated: Apr 13, 2026

Measuring Composition of CD95 Death-Inducing Signaling Complex and Processing of Procaspase-8 in this Complex

Measuring Composition of CD95 Death-Inducing Signaling Complex and Processing of Procaspase-8 in this Complex

Published on: August 2, 2021

Protein-fragment complementation assays to analyze the CD95 complex.

Dhouha Msalbi¹, Eden Lebrault¹, Patrick Legembre¹

¹INSERM U1262, CNRS UMR 7276, CRIBL, University of Limoges, Limoges, France.

Methods in Cell Biology

|April 11, 2026

Summary

This summary is machine-generated.

This study introduces a protein-fragment complementation assay (PCA) to identify protein interactions. PCA was used to validate the self-interaction of CD95/Fas and its interaction with FADD, crucial for immune homeostasis.

Keywords:

Apoptosis CD95 FADD Fas Inflammation Protein complementation assay

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Last Updated: Apr 13, 2026

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Published on: August 2, 2021

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Dissecting Multi-protein Signaling Complexes by Bimolecular Complementation Affinity Purification BiCAP

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Published on: June 15, 2018

Area of Science:

Immunology
Molecular Biology
Biochemistry

Background:

CD95/Fas receptor and its ligand CD95L are key in immune homeostasis.
CD95 has distinct domains (CRDs 1-3) involved in ligand binding and receptor aggregation.
CD95 signaling involves recruiting FADD and caspase-8 to form the death-inducing signaling complex (DISC).

Purpose of the Study:

To describe a novel protein-fragment complementation assay (PCA) for identifying protein-protein interactions (PPIs).
To validate the use of PCA for studying the interactions of CD95/Fas.
To confirm the self-interaction of CD95/Fas and its interaction with FADD.

Main Methods:

Protein-fragment complementation assay (PCA) utilizing a split Renilla reniformis luciferase (Rluc) reporter system.
Fusion of Rluc fragments to proteins of interest (POIs) to detect interactions.
Quantification of luciferase activity to indicate PPIs.

Main Results:

The PCA method was successfully established and utilized.
PCA validated the self-interaction of CD95/Fas.
PCA confirmed the interaction between CD95/Fas and FADD.

Conclusions:

Protein-fragment complementation assay (PCA) is an effective method for identifying and validating protein-protein interactions.
The findings support the role of CD95/Fas self-aggregation and FADD interaction in immune signaling pathways.
PCA offers a valuable tool for studying complex molecular interactions in biological systems.