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The riddle of mitochondrial caspase-3 from liver

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Abstract

Caspase-3 is one of the main executors of apoptosis. Its zymogen procaspase-3 was localized to cytosol, mitochondria and nuclei. The subcellular location of procaspase-3 in liver was reported by several studies to be either cytosolic or cytosolic and mitochondrial. Our aim was to investigate these separate procaspase-3 pools to differentiate the pathways of their activation. By cell fractionation, immunocytochemistry, and confocal microscopy we report that there is a single procaspase-3 pool located to the cytosol in primary hepatocytes and in fractions of rat liver. In contrast, it depends on the isolation purity whether procaspase-3 is located in mitochondria of non-parenchymal liver cells, or not. All preparations with mitochondrial procaspase-3 fractions contain traces of haemoglobin, indicating the presence of some erythrocytes, which are the source of mitochondrial procaspase-3. Since erythrocytes migrate with mitochondria in subcellular fractionations, it is important to check for haemoglobin, before localizing the protein to mitochondria.

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References

  1. Chandra D, Tang DG (2003) Mitochondrially localized active caspase-9 and caspase-3 result mostly from translocation from the cytosol and partly from caspase-mediated activation in the organelle. J Biol Chem 278:17408–17420

    Article  PubMed  CAS  Google Scholar 

  2. Samali A, Zhivotovsky B, Jones DP et al (1998) Detection of pro-caspase-3 in cytosol and mitochondria of various tissues. FEBS Lett 431:167–169

    Article  PubMed  CAS  Google Scholar 

  3. Mancini M, Nicholson DW, Roy S et al (1998) The caspase-3 precursor has a cytosolic and mitochondrial distribution: implications for apoptotic signaling. J Cell Biol 140:1485–1495

    Article  PubMed  CAS  Google Scholar 

  4. Ramuz O, Isnardon D, Devilard E et al (2003) Constitutive nuclear localization and initial cytoplasmic apoptotic activation of endogenous caspase-3 evidenced by confocal microscopy. Int J Exp Path 84:75–81

    Article  Google Scholar 

  5. Chandler JM, Cohen GM, MacFarlane M (1998) Different subcellular distribution of caspase-3 and caspase-7 following Fas-induced apoptosis in mouse liver. J Biol Chem 273:10815–10818

    Article  PubMed  CAS  Google Scholar 

  6. van Loo G, Saelens X, Matthijssens F et al (2002) Caspases are not localized in mitochondria during life or death. Cell Death Diff 9:1207–1211

    Article  Google Scholar 

  7. van Berkel TJC, Kruijt JK (1977) Different types of mitochondria in parenchymal and non-parenchymal rat-liver cells. Eur J Biochem 73:223–229

    Article  PubMed  Google Scholar 

  8. Krajewska M, Wang H-G, Krajewski S et al (1997) Immunohistochemical analysis of in vivo patterns of expression of CPP32 (caspase-3), a cell death protease. Cancer Res 57:1605–1613

    PubMed  CAS  Google Scholar 

  9. Berry MN, Friend DS (1969) High yield preparation of isolated rat liver parenchymal cells. J Cell Biol 48:506–520

    Article  Google Scholar 

  10. Werner S, Neupert W (1972) Functional and biogenetical heterogeneity of the inner membrane of rat-liver mitochondria. Eur J Biochem 25:379–396

    Article  PubMed  CAS  Google Scholar 

  11. Milisav I, Šuput D (2007) Procaspase-9 is attached to the mitochondrial outer membrane in the early stages of apoptosis. Cell Mol Biol Lett 12:509–522

    Article  PubMed  CAS  Google Scholar 

  12. Duan H, Orth K, Chinnaiyan AM et al (1996) ICE-LAP-6, a novel member of the ICE/Ced-3 gene family, is activated by the cytotoxic T cell protease granzyme B. J Biol Chem 271:16720–16724

    Article  PubMed  CAS  Google Scholar 

  13. Matocha MF, Waterman MR (1984) Discriminatory processing of the precursor forms of cytochrome P-450SCC and adrenodoxin by adrenocortical and heart mitochondria. J Biol Chem 259:8672–8678

    PubMed  CAS  Google Scholar 

  14. Pelham HR, Jackson RJ (1976) An efficient mRNA-dependent translation system from reticulocyte lysates. Eur J Biochem 67:247–256

    Article  PubMed  CAS  Google Scholar 

  15. Colin M, Moriz S, Schneider H et al (2000) Haemoglobin interferes with the ex vivo luciferase luminescence assay: consequence for detection of luciferase reporter gene expression in vivo. Gene Therapy 7:1333–1336

    Article  PubMed  CAS  Google Scholar 

  16. Castell JV, Jover R, Martinez-Jimenez CP et al (2006) Hepatocyte cell lines: their use, scope and limitations in drug metabolism studies. Expert Opin Drug Metab Toxicol 2:183–212

    Article  PubMed  CAS  Google Scholar 

  17. Okado-Matsumoto A, Fridovich I (2001) Subcellular distribution of superoxide dismutases (SOD) in rat liver. J Biol Chem 276:38388–38393

    Article  PubMed  CAS  Google Scholar 

  18. Geller BL, Winge DR (1982) Rat liver Cu, Zn-superoxide dismutase. J Biol Chem 257:8945–8952

    PubMed  CAS  Google Scholar 

  19. Berg CP, Engels IH, Rothbart A et al (2001) Human mature red blood cells express caspase-3 and caspase-8, but are devoid of mitochondrial regulators of apoptosis. Cell Death Diff 8:1197–1206

    Article  CAS  Google Scholar 

  20. Bratosin D, Estaquier J, Petit F et al (2001) Programmed cell death in mature erythrocytes: a model for investigating death effector pathways operating in the absence of mitochondria. Cell Death Diff 8:1143–1156

    Article  CAS  Google Scholar 

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Acknowledgments

We thank Dr. Rok Romih for assistance with immunoelectron microscopy, Dr. Helena Podgornik for advice on erythrocyte detection, Dr. Marko Živin and Andrej Vovk for advices on band densitometry and statistics, respectively. Alenka Frangež Štrukelj and Dejan Bijelič helped us with preparation of this manuscript. This work was supported by grant P3-0019 from Ministry of Higher Education, Science and Technology of Republic of Slovenia.

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Authors and Affiliations

  1. Institute of Pathophysiology, Faculty of Medicine, University of Ljubljana, Zaloška 4, 1000, Ljubljana, Slovenia

    Irina Milisav, Damijan Nipič & Dušan Šuput

  2. Faculty of Health Sciences, University of Ljubljana, Ljubljana, Slovenia

    Irina Milisav

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  1. Irina Milisav
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  2. Damijan Nipič
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  3. Dušan Šuput
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Correspondence to Irina Milisav.

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Milisav, I., Nipič, D. & Šuput, D. The riddle of mitochondrial caspase-3 from liver. Apoptosis 14, 1070–1075 (2009). https://doi.org/10.1007/s10495-009-0381-3

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