Apoptosis by a cytosolic extract from Fas-activated cells

EMBO J. 1995 Nov 1;14(21):5201-8.

Abstract

Fas is a type I membrane protein and its activation by binding of the Fas ligand or an agonistic anti-Fas antibody induces apoptosis in Fas-bearing cells. In this report we prepared lysates from cells treated with anti-Fas antibody. The lysates induced apoptotic morphological changes in nuclei from normal mouse liver, accompanied by DNA degradation. The apoptosis-inducing activity was quickly generated in cells by anti-Fas antibody and was found in the soluble cytosolic fraction. Induction of the activity in cells was inhibited by a tetrapeptide, acetyl-Tyr-Val-Ala-Asp-chloromethylketone, a specific inhibitor of interleukin-1 beta converting enzyme. Addition of COS cell lysates containing Bcl-2 to the assay significantly inhibited the apoptotic process, indicating that the in vitro process reflected apoptosis that occurs in intact cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Apoptosis / drug effects
  • Apoptosis / physiology*
  • Caspase 1
  • Cell Extracts
  • Cell Line
  • Cell Nucleus / physiology
  • Cysteine Endopeptidases / metabolism
  • Cysteine Proteinase Inhibitors / pharmacology
  • Cytosol / physiology
  • DNA Damage / drug effects
  • Liver / pathology
  • Mice
  • Proto-Oncogene Proteins / physiology
  • Proto-Oncogene Proteins c-bcl-2
  • fas Receptor / immunology
  • fas Receptor / physiology*

Substances

  • Cell Extracts
  • Cysteine Proteinase Inhibitors
  • Proto-Oncogene Proteins
  • Proto-Oncogene Proteins c-bcl-2
  • fas Receptor
  • Cysteine Endopeptidases
  • Caspase 1