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PMID: 9892620 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Mitochondrial release of caspase-2 and -9 during the apoptotic process.

The Journal of experimental medicine ·Vol. 189 ·No. 2 ·1999-01-18 ·Pages 381-94

Susin SA, Lorenzo HK, Zamzami N, Marzo I, Brenner C, Larochette N, Prévost MC, Alzari PM, Kroemer G

Abstract

The barrier function of mitochondrial membranes is perturbed early during the apoptotic process. Here we show that the mitochondria contain a caspase-like enzymatic activity cleaving the caspase substrate Z-VAD.afc, in addition to three biological activities previously suggested to participate in the apoptotic process: (a) cytochrome c; (b) an apoptosis-inducing factor (AIF) which causes isolated nuclei to undergo apoptosis in vitro; and (c) a DNAse activity. All of these factors, which are biochemically distinct, are released upon opening of the permeability transition (PT) pore in a coordinate, Bcl-2-inhibitable fashion. Caspase inhibitors fully neutralize the Z-VAD.afc-cleaving activity, have a limited effect on the AIF activity, and have no effect at all on the DNase activities. Purification of proteins reacting with the biotinylated caspase substrate Z-VAD, immunodetection, and immunodepletion experiments reveal the presence of procaspase-2 and -9 in mitochondria. Upon induction of PT pore opening, these procaspases are released from purified mitochondria and become activated. Similarly, upon induction of apoptosis, both procaspases redistribute from the mitochondrion to the cytosol and are processed to generate enzymatically active caspases. This redistribution is inhibited by Bcl-2. Recombinant caspase-2 and -9 suffice to provoke full-blown apoptosis upon microinjection into cells. Altogether, these data suggest that caspase-2 and -9 zymogens are essentially localized in mitochondria and that the disruption of the outer mitochondrial membrane occurring early during apoptosis may be critical for their subcellular redistribution and activation.

MeSH Terms
Animals Apoptosis/physiology Apoptosis Inducing Factor Caspase 2 Caspase 9 Caspases/metabolism Cell Line Cysteine Proteinase Inhibitors/pharmacology Cytochrome c Group/metabolism Enzyme Activation Enzyme Precursors/metabolism Female Flavoproteins Humans Intracellular Membranes/metabolism Membrane Proteins/metabolism Mice Mice, Inbred BALB C Microinjections Mitochondria/enzymology Nuclear Proteins/metabolism Proto-Oncogene Proteins c-bcl-2/metabolism Recombinant Proteins/metabolism
Chemicals
AIFM1 protein, human Apoptosis Inducing Factor Cysteine Proteinase Inhibitors Cytochrome c Group Enzyme Precursors Flavoproteins Membrane Proteins Nuclear Proteins Pdcd8 protein, mouse Proto-Oncogene Proteins c-bcl-2 Recombinant Proteins CASP9 protein, human Casp9 protein, mouse Caspase 2 Caspase 9 Caspases
Authors & Affiliations
9 authors, click to expand affiliations / ORCID
Susin S A
Centre National de la Recherche Scientifique, UPR 420, F-94801 Villejuif, France.
Lorenzo H K
Zamzami N
Marzo I
Brenner C
Larochette N
Prévost M C
Alzari P M
Kroemer G
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Article Info
Journal
The Journal of experimental medicine
Abbr.
J Exp Med
ISSN
0022-1007
DOI
10.1084/jem.189.2.381
Published
1999-01-18
Pages
381-94
Language
English
Region
United States
NLM ID
2985109R
PMCID
PMC2192979
Subset
IM
External Links
PubMed source DOI: 10.1084/jem.189.2.381 PMC Full Text View journal details
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