PMID- 20640600
OWN - NLM
STAT- MEDLINE
DCOM- 20110309
LR  - 20240109
IS  - 1573-675X (Electronic)
IS  - 1360-8185 (Linking)
VI  - 15
IP  - 12
DP  - 2010 Dec
TI  - Calpain and caspase processing of caspase-12 contribute to the ER stress-induced 
      cell death pathway in differentiated PC12 cells.
PG  - 1480-93
LID - 10.1007/s10495-010-0526-4 [doi]
AB  - Neuronal cell death after traumatic brain injury, Alzheimer's disease and 
      ischemic stroke may in part be mediated through endoplasmic reticulum (ER) stress 
      and unfolded protein response (UPR). UPR results in induction of molecular 
      chaperone GRP78 and the ER-resident caspase-12, whose activation has been 
      proposed to be mediated by calpain and caspase processing, although their 
      relative contribution remains unclear. In this study we induced ER stress with 
      thapsigargin (TG), and determined the activation profile of calpain-2, caspase-3, 
      caspase-7, and caspase-12 by analyses of protein levels, corresponding substrates 
      and breakdown products (BDP). Specific calpain and caspase activity was assessed 
      by analysis of alphaII-spectrin BDP of 145 kDa (SBDP145), BDP of 150 kDa (SBDP150) 
      and BDP of 120 kDa (SBDP120). Decrease in pro-calpain-2 protein and increased 
      SBDP145 levels by 3 h after TG treatment indicated early calpain activity. Active 
      caspase-7 (p20) increase occurred after 8 h, followed by concomitant 
      up-regulation of active caspase-3 and SBDP120 after 24 h. In vitro digestion 
      experiments supported that SBDP120 was exclusively generated by active caspase-3 
      and validated that kinectin and co-chaperone p23 were calpain and caspase-7 
      substrates, respectively. Pro-caspase-12 protein processing by the specific 
      action of calpain and caspase-3/7 was observed in a time-dependent manner. 
      N-terminal pro-domain processing of pro-caspase-12 by calpain generated a 38 kDa 
      fragment, while caspase-3/7 generated a 35 kDa fragment. Antibody developed 
      specifically against the caspase-3/7 C-terminal cleavage site D(341) detected the 
      presence of large subunit (p20) containing 23 kDa fragment that increased after 
      24 h of TG treatment. Significant caspase-12 enzyme activity was only detected 
      after 24 h of TG treatment and was completely inhibited by caspase 3/7 inhibitor 
      DEVD-fmk and partially by calpain inhibitor SNJ-1945. ER-stress-induced cell 
      death pathway in TG-treated PC12 cells was characterized by up-regulation of 
      GRP-78 and processing and activation of caspase-12 by the orchestrated 
      proteolytic activity of calpain-2 and caspase-3/7.
FAU - Martinez, Juan A
AU  - Martinez JA
AD  - Center of Innovative Research, Banyan Biomarkers Inc, 12085 Research Drive, 
      Alachua, FL 32615, USA. jmartinez@banyanbio.com
FAU - Zhang, Zhiqun
AU  - Zhang Z
FAU - Svetlov, Stanislav I
AU  - Svetlov SI
FAU - Hayes, Ronald L
AU  - Hayes RL
FAU - Wang, Kevin K
AU  - Wang KK
FAU - Larner, Stephen F
AU  - Larner SF
LA  - eng
GR  - R01NS051431/NS/NINDS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PL  - Netherlands
TA  - Apoptosis
JT  - Apoptosis : an international journal on programmed cell death
JID - 9712129
RN  - 0 (Endoplasmic Reticulum Chaperone BiP)
RN  - 0 (Enzyme Inhibitors)
RN  - 0 (HSPA5 protein, human)
RN  - 0 (Heat-Shock Proteins)
RN  - 0 (Microfilament Proteins)
RN  - 0 (Peptide Fragments)
RN  - 0 (Sptan1 protein, rat)
RN  - 0 (Vesicular Transport Proteins)
RN  - 67526-95-8 (Thapsigargin)
RN  - EC 3.4.22.- (CASP12 protein, human)
RN  - EC 3.4.22.- (Calpain)
RN  - EC 3.4.22.- (Caspase 12)
RN  - EC 3.4.22.- (Caspases)
SB  - IM
MH  - Amino Acid Sequence
MH  - Animals
MH  - *Calpain/metabolism
MH  - Caspase 12/*metabolism
MH  - *Caspases/metabolism
MH  - Cell Death/drug effects/*physiology
MH  - Cell Differentiation
MH  - Cell Line, Tumor
MH  - Endoplasmic Reticulum/drug effects/*enzymology
MH  - Endoplasmic Reticulum Chaperone BiP
MH  - Enzyme Activation/drug effects/*physiology
MH  - Enzyme Inhibitors/pharmacology
MH  - Heat-Shock Proteins/*metabolism
MH  - Humans
MH  - Microfilament Proteins/*analysis/chemistry
MH  - Molecular Sequence Data
MH  - Neurons/chemistry/physiology
MH  - Oxidative Stress/drug effects/*physiology
MH  - Peptide Fragments/*analysis/chemistry
MH  - Rats
MH  - Substrate Specificity
MH  - Thapsigargin/pharmacology
MH  - Time Factors
MH  - Unfolded Protein Response/drug effects/*physiology
MH  - Vesicular Transport Proteins/*analysis/chemistry
EDAT- 2010/07/20 06:00
MHDA- 2011/03/10 06:00
CRDT- 2010/07/20 06:00
PHST- 2010/07/20 06:00 [entrez]
PHST- 2010/07/20 06:00 [pubmed]
PHST- 2011/03/10 06:00 [medline]
AID - 10.1007/s10495-010-0526-4 [doi]
PST - ppublish
SO  - Apoptosis. 2010 Dec;15(12):1480-93. doi: 10.1007/s10495-010-0526-4.