Document Detail


Caspase-dependent activation of calpain during drug-induced apoptosis.
MedLine Citation:
PMID:  10075737     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
We have previously demonstrated that calpain is responsible for the cleavage of Bax, a proapoptotic protein, during drug-induced apoptosis of HL-60 cells (Wood, D. E., Thomas, A., Devi, L. A., Berman, Y., Beavis, R. C., Reed, J. C., and Newcomb, E. W. (1998) Oncogene 17, 1069-1078). Here we show the sequential activation of caspases and calpain during drug-induced apoptosis of HL-60 cells. Time course experiments using the topoisomerase I inhibitor 9-amino-20(S)-camptothecin revealed that cleavage of caspase-3 substrates poly(ADP-ribose) polymerase (PARP) and the retinoblastoma protein as well as DNA fragmentation occurred several hours before calpain activation and Bax cleavage. Pretreatment with the calpain inhibitor calpeptin blocked calpain activation and Bax cleavage but did not inhibit PARP cleavage, DNA fragmentation, or 9-amino-20(S)-camptothecin-induced morphological changes and cell death. Pretreatment with the pan-caspase inhibitor benzyloxycarbonyl-Val-Ala-Asp-fluoromethylketone (Z-VAD-fmk) inhibited PARP cleavage, DNA fragmentation, calpain activation, and Bax cleavage and increased cell survival by 40%. Interestingly, Z-VAD-fmk-treated cells died in a caspase- and calpain-independent manner that appeared morphologically distinct from apoptosis. Our results suggest that excessive or uncontrolled calpain activity may play a role downstream of and distinct from caspases in the degradation phase of apoptosis.
Authors:
D E Wood; E W Newcomb
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  The Journal of biological chemistry     Volume:  274     ISSN:  0021-9258     ISO Abbreviation:  J. Biol. Chem.     Publication Date:  1999 Mar 
Date Detail:
Created Date:  1999-04-15     Completed Date:  1999-04-15     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  2985121R     Medline TA:  J Biol Chem     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  8309-15     Citation Subset:  IM    
Affiliation:
Department of Pathology, New York University Medical Center, New York, New York 10016, USA.
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MeSH Terms
Descriptor/Qualifier:
Amino Acid Chloromethyl Ketones / pharmacology
Apoptosis* / drug effects
Calpain / metabolism*
Caspase 3
Caspases / metabolism*
Coumarins / pharmacology
Cysteine Proteinase Inhibitors / pharmacology
DNA Fragmentation
Dipeptides / pharmacology
Enzyme Activation
HL-60 Cells
Humans
Oligopeptides / pharmacology
Chemical
Reg. No./Substance:
0/Ac-aspartyl-glutamyl-valyl-aspartyl-aminomethylcoumarin; 0/Amino Acid Chloromethyl Ketones; 0/Coumarins; 0/Cysteine Proteinase Inhibitors; 0/Dipeptides; 0/Oligopeptides; 0/acetyl-aspartyl-glutamyl-valyl-aspartal; 0/benzyloxycarbonylvalyl-alanyl-aspartyl fluoromethyl ketone; 117591-20-5/calpeptin; EC 3.4.22.-/CASP3 protein, human; EC 3.4.22.-/Calpain; EC 3.4.22.-/Caspase 3; EC 3.4.22.-/Caspases

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