Roy, Ankoor Hong, Jong hui Lee, Jin-Hee Lee, Young-Tae Lee, Bong-Jin Kim, Key-Sun 2024-01-20T23:01:36Z 2024-01-20T23:01:36Z 2021-09-03 2008-08 1016-8478 https://pubs.kist.re.kr/handle/201004/133290 Procaspase-8 is activated by forming a death-inducing signaling complex (DISC) with the Fas-associated death domain (FADD) and the Fas receptor, but the mechanism of its activation is not well understood. Procaspase-8 devoid of the death effector domain at its N-terminus (Delta nprocaspase-8) was reported to be activated by kosmotropic salts, but it has not been induced to form a DISC in vitro because it cannot interact with FADD. Here, we report the production of full-length procaspase-8 and show that it is activated by adding the Fas death domain (Fas-DD) and the FADD forming the cytoplasmic part of the DISC (cDISC). Furthermore, mutations known to affect DISC formation in vivo were shown to have the same effect on procaspase-8 activation in vitro. An antibody that induces Fas-DD association enhanced procaspase-8 activation, suggesting that the Fas ligand is not required for low-level activation of procaspase-8, but that Fas receptor clustering is needed for high-level activation of procaspase-8 leading to cell death. In vitro activation of procaspase-8 by forming a cDISC will be invaluable for investigating activation of ligand-mediated apoptosis and the numerous interactions affecting procaspase-8 activation. English KOREAN SOC MOLECULAR & CELLULAR BIOLOGY In vitro activation of procaspase-8 by forming the cytoplasmic component of the death-inducing signaling complex (cDISC) Article 1 MOLECULES AND CELLS, v.26, no.2, pp.165 - 170 MOLECULES AND CELLS 26 2 165 170 N scie scopus kci ART001274470 000258949800009 2-s2.0-53749091594 Biochemistry & Molecular Biology Cell Biology Biochemistry & Molecular Biology Cell Biology Article CELL-DEATH FAS CASPASE-8 DOMAIN FADD OLIGOMERIZATION PROTEIN MODEL FLICE DISC in vitro activation procaspase-8