Document Detail


Calpain-mediated Bid cleavage and calpain-independent Bak modulation: two separate pathways in cisplatin-induced apoptosis.
MedLine Citation:
PMID:  11940658     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Calpain is a ubiquitous protease with potential involvement in apoptosis. We report that in human melanoma cells, cisplatin-induced calpain activation occurs early in apoptosis. Calpain activation and subsequent apoptosis were inhibited by calpeptin and PD150606, two calpain inhibitors with different modes of action. Furthermore, cisplatin induced cleavage of the BH3-only protein Bid, yielding a 14-kDa fragment similar to proapoptotic, caspase-cleaved Bid. However, Bid cleavage was inhibited by inhibitors of calpain, but not by inhibitors of caspases or of cathepsin L. Recombinant Bid was cleaved in vitro by both recombinant calpain and by lysates of cisplatin-treated cells. Cleavage was calpeptin sensitive, and the cleavage site was mapped between Gly70 and Arg71. Calpain-cleaved Bid induced cytochrome c release from isolated mitochondria. While calpeptin did not affect cisplatin-induced modulation of Bak to its proapoptotic conformation, a dominant-negative mutant of MEKK1 (dnMEKK) inhibited Bak modulation. dnMEKK did not, however, block Bid cleavage. The combination of dnMEKK and calpeptin had an additive inhibitory effect on apoptosis. In summary, calpain-mediated Bid cleavage is important in drug-induced apoptosis, and cisplatin induces at least two separate apoptotic signaling pathways resulting in Bid cleavage and Bak modulation, respectively.
Authors:
Aleksandra Mandic; Kristina Viktorsson; Linda Strandberg; Thomas Heiden; Johan Hansson; Stig Linder; Maria C Shoshan
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Molecular and cellular biology     Volume:  22     ISSN:  0270-7306     ISO Abbreviation:  Mol. Cell. Biol.     Publication Date:  2002 May 
Date Detail:
Created Date:  2002-04-09     Completed Date:  2002-05-16     Revised Date:  2009-11-19    
Medline Journal Info:
Nlm Unique ID:  8109087     Medline TA:  Mol Cell Biol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  3003-13     Citation Subset:  IM    
Affiliation:
Cancer Center Karolinska, Department of Oncology-Pathology, Karolinska Institute and Hospital, S-171 76 Stockholm, Sweden.
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MeSH Terms
Descriptor/Qualifier:
Amino Acid Sequence
Apoptosis / drug effects*
Arginine / metabolism
BH3 Interacting Domain Death Agonist Protein
Calcium / metabolism
Calpain / antagonists & inhibitors,  metabolism*
Carrier Proteins / chemistry,  metabolism*
Cathepsin L
Cathepsins / antagonists & inhibitors,  metabolism
Cisplatin / pharmacology*
Cysteine Endopeptidases
Cytochrome c Group / metabolism
Dipeptides / pharmacology
Enzyme Activation
Glycine / metabolism
Humans
Intracellular Membranes / metabolism
Membrane Potentials / drug effects
Membrane Proteins / metabolism*
Mitochondria / metabolism
Molecular Sequence Data
Protein Processing, Post-Translational / drug effects*
Signal Transduction
Time Factors
Tumor Cells, Cultured
bcl-2 Homologous Antagonist-Killer Protein
Chemical
Reg. No./Substance:
0/BAK1 protein, human; 0/BH3 Interacting Domain Death Agonist Protein; 0/BID protein, human; 0/Carrier Proteins; 0/Cytochrome c Group; 0/Dipeptides; 0/Membrane Proteins; 0/bcl-2 Homologous Antagonist-Killer Protein; 117591-20-5/calpeptin; 15663-27-1/Cisplatin; 56-40-6/Glycine; 74-79-3/Arginine; 7440-70-2/Calcium; EC 3.4.-/Cathepsins; EC 3.4.22.-/Calpain; EC 3.4.22.-/Cysteine Endopeptidases; EC 3.4.22.15/CTSL1 protein, human; EC 3.4.22.15/Cathepsin L
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