Artikel: Measuring composition of cd95 death-inducing signaling complex and processing of procaspase-8 in this complex
Journal of visualized experiments. 2021 Aug. 02, , no. 174
2021
Abstract: Extrinsic apoptosis is mediated by the activation of death receptors (DRs) such as CD95/Fas/APO-1 or tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-receptor 1/receptor 2 (TRAIL-R1/R2). Stimulation of these receptors with their cognate ... ...
Abstract | Extrinsic apoptosis is mediated by the activation of death receptors (DRs) such as CD95/Fas/APO-1 or tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-receptor 1/receptor 2 (TRAIL-R1/R2). Stimulation of these receptors with their cognate ligands leads to the assembly of the death-inducing signaling complex (DISC). DISC comprises DR, the adaptor protein Fas-associated protein with death domain (FADD), procaspases-8/-10, and cellular FADD-like interleukin (IL)-1β-converting enzyme-inhibitory proteins (c-FLIPs). The DISC serves as a platform for procaspase-8 processing and activation. The latter occurs via its dimerization/oligomerization in the death effector domain (DED) filaments assembled at the DISC. Activation of procaspase-8 is followed by its processing, which occurs in several steps. In this work, an established experimental workflow is described that allows the measurement of DISC formation and the processing of procaspase-8 in this complex. The workflow is based on immunoprecipitation techniques supported by western blot analysis. This workflow allows careful monitoring of different steps of procaspase-8 recruitment to the DISC and its processing and is highly relevant for investigating molecular mechanisms of extrinsic apoptosis. |
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Schlagwörter | Western blotting ; apoptosis ; death ; dimerization ; interleukins ; ligands ; necrosis ; neoplasms ; oligomerization ; precipitin tests ; protein domains |
Sprache | Englisch |
Erscheinungsverlauf | 2021-0802 |
Umfang | p. e62842. |
Erscheinungsort | Journal of Visualized Experiments |
Dokumenttyp | Artikel |
ZDB-ID | 2259946-0 |
ISSN | 1940-087X |
ISSN | 1940-087X |
DOI | 10.3791/62842 |
Datenquelle | NAL Katalog (AGRICOLA) |
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