Abstract
Caspases are crucial for the initiation, propagation and execution of apoptosis. They normally exist as proenzymes, which can be activated through recruitment into activating complexes and by proteolytic cleavage by other caspases or proteases. Perturbation of organelles such as nuclei, endoplasmatic reticulum and mitochondria results in the activation of caspases. A number of caspases (-2, -3, -8 and -9) were published as being localized in the intermembrane space of mitochondria. However, in three different models of apoptosis (anti-Fas-induced cell death in murine hepatocytes, Fas ligand-induced apoptosis in Jurkat cells and apoptosis induced by growth factor withdrawal in Ba/F3 cells) we could not identify a mitochondrial location of caspases, neither under control nor under apoptotic conditions. In all three apoptotic models caspases were found in the cytosolic (caspases-2, -3, -6, -7, -8, -9) and nuclear subcellular fractions (caspases-2, -3). In another approach we treated isolated liver mitochondria with truncated Bid. Although tBid-dependent release of Cytochrome c, AIF, adenylate kinase, Smac/DIABLO and Omi/HtrA2 could be demonstrated, none of the caspases were detectable both in the supernatant and the mitochondrial fraction after treatment. Our results demonstrate that, in contrast to previous studies, no caspases-2, -3, -8 and -9 are associated with the mitochondrial fraction. These findings support the concept of a separate compartmentalization between proapoptotic cofactors in the mitochondria and silent precursor caspases in the cytosol.
Similar content being viewed by others
Log in or create a free account to read this content
Gain free access to this article, as well as selected content from this journal and more on nature.com
or
Abbreviations
- COX:
-
cytochrome oxidase
- Ac-DEVD-amc:
-
acetyl-Asp(Ome)-Glu(Ome)-Val-Asp(Ome)-aminomethylcoumarine
- tBid:
-
truncated Bid
References
Earnshaw WC, Martins LM, Kaufmann SH . 1999 Mammalian caspases: structure, activation, substrates, and functions during apoptosis Annu. Rev. Biochem. 68: 383–424
Lamkanfi M, Declercq W, Kalai M, Saelens X, Vandenabeele P . 2002 Alice in caspase land: a phylogenetic analysis of caspases from worm to man Cell. Death Differ 9 358–361
Van de Craen M, Vandenabeele P, Declercq W, Van den Brande I, Van Loo G, Molemans F, Schotte P, Van Criekinge W, Beyaert R, Fiers W . 1997 Characterization of seven murine caspase family members FEBS Lett. 403: 61–69
Van de Craen M, Van Loo G, Pype S, Van Criekinge W, Van den brande I, Molemans F, Fiers W, Declercq W, Vandenabeele P . 1998 Identification of a new caspase homologue: caspase-14 Cell. Death Differ. 5: 838–846
Van de Craen M, Van Loo G, Declercq W, Schotte P, Van den brande I, Mandruzzato S, van der Bruggen P, Fiers W, Vandenabeele P . 1998 Molecular cloning and identification of murine caspase-8 J. Mol. Biol. 284: 1017–1026
Lippens S, Kockx M, Knaapen M, Mortier L, Polakowska R, Verheyen A, Garmyn M, Zwijsen A, Formstecher P, Huylebroeck D, Vandenabeele P, Declercq W . 2000 Epidermal differentiation does not involve the pro-apoptotic executioner caspases, but is associated with caspase-14 induction and processing Cell. Death Differ. 7: 1218–1224
Mancini M, Nicholson DW, Roy S, Thornberry NA, Peterson EP, Casciola-Rosen LA, Rosen A . 1998 The caspase-3 precursor has a cytosolic and mitochondrial distribution: implications for apoptotic signaling J. Cell. Biol. 140: 1485–1495
Samali A, Zhivotovsky B, Jones DP, Orrenius S . 1998 Detection of pro-caspase-3 in cytosol and mitochondria of various tissues FEBS Lett. 431: 167–169
Krajewski S, Krajewska M, Ellerby LM, Welsh K, Xie Z, Deveraux QL, Salvesen GS, Bredesen DE, Rosenthal RE, Fiskum G, Reed JC . 1999 Release of caspase-9 from mitochondria during neuronal apoptosis and cerebral ischemia Proc. Natl. Acad. Sci. USA 96: 5752–5757
Susin SA, Lorenzo HK, Zamzami N, Marzo I, Brenner C, Larochette N, Prevost MC, Alzari PM, Kroemer G . 1999 Mitochondrial release of caspase-2 and -9 during the apoptotic process J. Exp. Med. 189: 381–394
Zhivotovsky B, Samali A, Gahm A, Orrenius S . 1999 Caspases: their intracellular localization and translocation during apoptosis Cell. Death Differ. 6: 644–651
Qin ZH, Wang Y, Kikly KK, Sapp E, Kegel KB, Aronin N, DiFiglia M . 2001 Pro-caspase-8 is predominantly localized in mitochondria and released into cytoplasm upon apoptotic stimulation J. Biol. Chem. 276: 8079–8086
Costantini P, Bruey JM, Castedo M, Metivier D, Loeffler M, Susin SA, Ravagnan L, Zamzami N, Garrido C, Kroemer G . 2002 Pre-processed caspase-9 contained in mitochondria participates in apoptosis Cell. Death Differ. 9: 82–88
Colussi PA, Harvey NL, Kumar S . 1998 Prodomain-dependent nuclear localization of the caspase-2 (Nedd2) precursor. A novel function for a caspase prodomain J. Biol. Chem. 273: 24535–24542
Mancini M, Machamer CE, Roy S, Nicholson DW, Thornberry NA, Casciola-Rosen LA, Rosen A . 2000 Caspase-2 is localized at the Golgi complex and cleaves golgin-160 during apoptosis J. Cell. Biol. 149: 603–612
Shikama Y, Mami U, Miyashita T, Yamada M . 2001 Comprehensive studies on subcellular localizations and cell death- inducing activities of eight GFP-tagged apoptosis-related caspases Exp. Cell. Res. 264: 315–325
Goldstein JC, Waterhouse NJ, Juin P, Evan GI, Green DR . 2000 The coordinate release of cytochrome c during apoptosis is rapid, complete and kinetically invariant Nat. Cell. Biol. 2: 156–162
Bergeron L, Perez GI, Macdonald G, Shi L, Sun Y, Jurisicova A, Varmuza S, Latham KE, Flaws JA, Salter JC, Hara H, Moskowitz MA, Li E, Greenberg A, Tilly JL, Yuan J . 1998 Defects in regulation of apoptosis in caspase-2-deficient mice Genes Dev. 12: 1304–1314
Leonhard K, Guiard B, Pellecchia G, Tzagoloff A, Neupert W, Langer T . 2000 Membrane protein degradation by AAA proteases in mitochondria: extraction of substrates from either membrane surface Mol. Cell. 5: 629–638
Kilic M, Schafer R, Hoppe J, Kagerhuber U . 2002 Formation of noncanonical high molecular weight caspase-3 and -6 complexes and activation of caspase-12 during serum starvation induced apoptosis in AKR-2B mouse fibroblasts Cell. Death Differ. 9: 125–137
Van Loo G, Demol H, van Gurp M, Hoorelbeke B, Schotte P, Beyaert R, Zhivotovsky B, Gevaert K, Declercq W, Vandekerckhove J, Vandenabeele P . 2002 A matrix-assisted laser desorption ionization post-source decay (MALDI- PSD) analysis of proteins released from isolated liver mitochondria treated with recombinant truncated Bid Cell. Death Differ. 9: 301–308
Van de Craen M, Declercq W, Van den brande I, Fiers W, Vandenabeele P . 1999 The proteolytic procaspase activation network: an in vitro analysis Cell Death Differ. 6: 1117–1124
Acknowledgements
The authors thank Dr. Michel Hahne (Centro Nacional de Biotecnologia, Madrid, Spain), Dr. Sharad Kumar (Hanson Centre for Cancer Research, Adelaide, Australia), Dr. Emad Alnemri (Kimmel Cancer Institute, Thomas Jefferson University, Philadelphia, Pennsylvania, USA), Dr. Adolf Ruiz-Carrillo (Department of Molecular and Cell Biology, C.I.D., C.S.I.C, Barcelona, Spain), Dr. Boris Zhivotovsky (Karolinska Institute, Institute of Environmental Medicine, Stockholm, Sweden) and Dr. Patrice Petit (Institut Cochin de Génétique Moléculaire, Paris, France) for providing a plasmid expressing soluble FasL, anti-caspase 2 antibody, anti-Omi/HtrA2, anti-endonuclease G, anti-AK2 and anti-AIF, respectively. We thank Ann Meeuws and Wilma Burm for expert technical assistance and Myriam Goessens and Leen Puimege for animal care. This work was supported in part by the Interuniversitaire Attractiepolen V, the Fonds voor Wetenschappelijk Onderzoek–Vlaanderen (grant 3G.0006.01 and grant 3G.021199), an EC-RTD grant QLG1-CT-1999-00739, a RUG-cofinanciering EU project (011C0300), and a RUG-GOA project (12050502).
Author information
Authors and Affiliations
Corresponding author
Additional information
Edited by S Kumar
Rights and permissions
About this article
Cite this article
van Loo, G., Saelens, X., Matthijssens, F. et al. Caspases are not localized in mitochondria during life or death. Cell Death Differ 9, 1207–1211 (2002). https://doi.org/10.1038/sj.cdd.4401101
Received:
Revised:
Accepted:
Published:
Issue date:
DOI: https://doi.org/10.1038/sj.cdd.4401101
Keywords
This article is cited by
-
Taxifolin, a natural flavonoid interacts with cell cycle regulators causes cell cycle arrest and causes tumor regression by activating Wnt/ β -catenin signaling pathway
BMC Cancer (2018)
-
Apoptosis regulation by subcellular relocation of caspases
Scientific Reports (2018)
-
More alive than dead: non-apoptotic roles for caspases in neuronal development, plasticity and disease
Cell Death & Differentiation (2017)
-
Caspase-2 resides in the mitochondria and mediates apoptosis directly from the mitochondrial compartment
Cell Death Discovery (2016)
-
Two-Photon Enzymatic Probes Visualizing Sub-cellular/Deep-brain Caspase Activities in Neurodegenerative Models
Scientific Reports (2016)
