Differential affinity of FLIP and procaspase 8 for FADD's DED binding surfaces regulates DISC assembly

Nat Commun. 2014 Feb 28:5:3350. doi: 10.1038/ncomms4350.

Abstract

Death receptor activation triggers recruitment of FADD, which via its death effector domain (DED) engages the DEDs of procaspase 8 and its inhibitor FLIP to form death-inducing signalling complexes (DISCs). The DEDs of FADD, FLIP and procaspase 8 interact with one another using two binding surfaces defined by α1/α4 and α2/α5 helices, respectively. Here we report that FLIP has preferential affinity for the α1/α4 surface of FADD, whereas procaspase 8 has preferential affinity for FADD's α2/α5 surface. These relative affinities contribute to FLIP being recruited to the DISC at comparable levels to procaspase 8 despite lower cellular expression. Additional studies, including assessment of DISC stoichiometry and functional assays, suggest that following death receptor recruitment, the FADD DED preferentially engages FLIP using its α1/α4 surface and procaspase 8 using its α2/α5 surface; these tripartite intermediates then interact via the α1/α4 surface of FLIP DED1 and the α2/α5 surface of procaspase 8 DED2.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Blotting, Western
  • CASP8 and FADD-Like Apoptosis Regulating Protein / metabolism*
  • Caspase 8 / metabolism*
  • Chromatography, Gel
  • Fas-Associated Death Domain Protein / metabolism*
  • HCT116 Cells
  • Humans
  • Immunoprecipitation
  • Protein Binding

Substances

  • CASP8 and FADD-Like Apoptosis Regulating Protein
  • FADD protein, human
  • Fas-Associated Death Domain Protein
  • Caspase 8