Sequential activation of ICE-like and CPP32-like proteases during Fas-mediated apoptosis

Nature. 1996 Apr 25;380(6576):723-6. doi: 10.1038/380723a0.


Binding of Fas ligand or an agonistic anti-Fas antibody induces apoptosis in Fas-bearing cells. The interleukin-1Beta-converting enzyme (ICE) is a cysteine protease that is involved in apoptosis induced by various stimuli, including Fas-mediated apoptosis. Several ICE homologues have been identified, and these are subdivided into three groups (ICE-, CPP32-, and Ich-1-like proteases). We show here that specific inhibitors of ICE- or CPP32-like proteases can inhibit Fas-mediated apoptosis. Transient ICE-like activity was found in the cytosolic fraction of Fas-activated cells, whereas ICE-dependent, CPP32-like activity gradually accumulated in the cytosol. Cell lysates from mouse lymphoma supplemented with either recombinant ICE or CPP32 induced apoptosis of nuclei. The CPP32 inhibitor inhibited ICE- or CPP32-induced apoptosis in the cell-free system, whereas the ICE-inhibitor only inhibited ICE-induced apoptosis. Cell extracts from thymocytes from ICE-null mice induced apoptosis in the cell-free system when it was supplemented with CPP32. These results indicate that Fas sequentially activates ICE- and CPP32-like proteases, and that downstream CPP32, together with a component(s) in the cytoplasm, causes apoptosis of nuclei.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Apoptosis / drug effects
  • Apoptosis / physiology*
  • Caspase 1
  • Caspase 3
  • Caspases*
  • Cell Line, Transformed
  • Cell Nucleus / enzymology
  • Cell Nucleus / physiology
  • Cysteine Endopeptidases / chemistry
  • Cysteine Endopeptidases / metabolism*
  • Cysteine Proteinase Inhibitors / pharmacology
  • Cytosol / enzymology
  • Enzyme Activation
  • Mice
  • Molecular Sequence Data
  • Oligopeptides / pharmacology
  • Peptides / metabolism
  • Recombinant Proteins / pharmacology
  • fas Receptor / physiology*


  • Cysteine Proteinase Inhibitors
  • Oligopeptides
  • Peptides
  • Recombinant Proteins
  • acetyl-aspartyl-glutamyl-valyl-aspartal
  • fas Receptor
  • L 709049
  • Casp3 protein, mouse
  • Caspase 3
  • Caspases
  • Cysteine Endopeptidases
  • Caspase 1