Stoichiometry of the CD95 Death-Inducing Signaling Complex: Experimental and Modeling Evidence for a Death Effector Domain Chain Model

The CD95 (Fas/APO-1) death-inducing signaling complex (DISC) is essential for the initiation of CD95-mediated apoptotic and nonapoptotic responses. The CD95 DISC comprises CD95, FADD, procaspase-8, procaspase-10, and c-FLIP proteins. Procaspase-8 and procaspase-10 are activated at the DISC, leading...

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Bibliographic Details
Published in:Molecular cell 2012-07, Vol.47 (2), p.306-319
Main Authors: Schleich, Kolja, Warnken, Uwe, Fricker, Nicolai, Öztürk, Selcen, Richter, Petra, Kammerer, Kerstin, Schnölzer, Martina, Krammer, Peter H., Lavrik, Inna N.
Format: Article
Language:English
Subjects:
Apoptosis
Caspase 10 - metabolism
Caspase 8 - metabolism
caspase-8
death
Death Domain Receptor Signaling Adaptor Proteins - metabolism
Dimerization
fas Receptor - chemistry
fas Receptor - metabolism
Fas-Associated Death Domain Protein - metabolism
HeLa Cells
Humans
mass spectrometry
Mass Spectrometry - methods
mathematical models
Microscopy, Fluorescence - methods
Models, Biological
Models, Theoretical
proteins
Signal Transduction
stoichiometry
Western blotting
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Summary:The CD95 (Fas/APO-1) death-inducing signaling complex (DISC) is essential for the initiation of CD95-mediated apoptotic and nonapoptotic responses. The CD95 DISC comprises CD95, FADD, procaspase-8, procaspase-10, and c-FLIP proteins. Procaspase-8 and procaspase-10 are activated at the DISC, leading to the formation of active caspases and apoptosis initiation. In this study we analyzed the stoichiometry of the CD95 DISC. Using quantitative western blots, mass spectrometry, and mathematical modeling, we reveal that the amount of DED proteins procaspase-8/procaspase-10 and c-FLIP at the DISC exceeds that of FADD by several-fold. Furthermore, our findings imply that procaspase-8, procaspase-10, and c-FLIP could form DED chains at the DISC, enabling the formation of dimers and efficient activation of caspase-8. Taken together, our findings provide an enhanced understanding of caspase-8 activation and initiation of apoptosis at the DISC. [Display omitted] ► The amount of procaspase-8/procaspase-10 and c-FLIP at the DISC exceeds that of FADD ► Procaspase-8, procaspase-10, and c-FLIP could form DED chains/platforms at the DISC, ► The length of the DED chains is variable and depends on the stimulation strength ► Mathematical model supported by quantitative mass spectrometry and western blot
ISSN:1097-2765
1097-4164
DOI:10.1016/j.molcel.2012.05.006