Processing of the Nedd2 precursor by ICE-like proteases and granzyme B

Genes Cells. 1996 Jul;1(7):673-85. doi: 10.1046/j.1365-2443.1996.00255.x.


Background: The Nedd2/Ich-1 protein belongs to a growing family of mammalian cysteine proteases similar to interleukin-1 beta converting enzyme (ICE). Because of their similarity to the Cacnorhabditis elegans cell death protein CED-3, the ICE-like proteins are thought to play a key role in the execution of apoptosis. The active form of ICE is a tetramer consisting of two heterodimers (p20 + p10)2 derived from the cleavage of the pro-enzyme.

Results: In the present communication we show that the p51 Nedd2 precursor (pro-Nedd2) is also cleaved into p20-like (p19) and p10-like (p12) subunits by extracts prepared from cultured cell lines. Extracts from apoptotic NIH-3T3 cells but not normal growing NIH-3T3 cells also contained pro-Nedd2 cleaving activity. The processing of pro-Nedd2 by cell extracts was inhibited by characteristic inhibitors of ICE-like proteases. Additionally we show that pro-Nedd2 (p51) can be processed in vitro by active CPP32 and ICE, and to a lesser extent by Mch2 and Nedd2. Granzyme B, a serine protease required for cytotoxic T lymphocyte (CTL) mediated killing of target cells, also cleaved pro-Nedd2 to p19 + p12 subunits.

Conclusions: Our observations suggest that Nedd2 activation requires cleavage by one or more ICE-like proteases that lie upstream in the proteolytic cascade. Cleavage of pro-Nedd2 by granzyme B indicates that Nedd2 may be one of the downstream effectors in the CTL-mediated killing of target cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3T3 Cells
  • Animals
  • Apoptosis
  • Base Sequence
  • Caenorhabditis elegans
  • Caspase 1
  • Caspase 2
  • Cysteine Endopeptidases / chemistry
  • Cysteine Endopeptidases / genetics
  • Cysteine Endopeptidases / metabolism*
  • DNA Primers / genetics
  • Enzyme Precursors / chemistry
  • Enzyme Precursors / genetics
  • Enzyme Precursors / metabolism*
  • Granzymes
  • Humans
  • In Vitro Techniques
  • Mice
  • Protease Inhibitors / pharmacology
  • Protein Conformation
  • Protein Processing, Post-Translational
  • Recombinant Proteins / metabolism
  • Serine Endopeptidases / metabolism*
  • T-Lymphocytes, Cytotoxic / metabolism


  • DNA Primers
  • Enzyme Precursors
  • Protease Inhibitors
  • Recombinant Proteins
  • GZMB protein, human
  • Granzymes
  • Gzmb protein, mouse
  • Serine Endopeptidases
  • CASP2 protein, human
  • Casp2 protein, mouse
  • Caspase 2
  • Cysteine Endopeptidases
  • Caspase 1