Molecules and Cells

Korean Society for Molecular and Cellular Biology (한국분자세포생물학회)
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In vitro Activation of Procaspase-8 by Forming the Cytoplasmic Component of the Death-inducing Signaling Complex (cDISC)

  • Roy, Ankoor (Center for Neural Science, Korea Institute of Science and Technology) ;
  • Hong, Jong hui (Center for Neural Science, Korea Institute of Science and Technology) ;
  • Lee, Jin-Hee (Center for Neural Science, Korea Institute of Science and Technology) ;
  • Lee, Young-Tae (Center for Neural Science, Korea Institute of Science and Technology) ;
  • Lee, Bong-Jin (Department of Pharmacy, Seoul National University) ;
  • Kim, Key-Sun (Center for Neural Science, Korea Institute of Science and Technology)
  • Received : 2008.01.04
  • Accepted : 2008.03.21
  • Published : 2008.08.31
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Abstract

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.

Keywords

Acknowledgement

Supported by : Korean Ministry of Science and Technology

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