Procaspase-3 and poly(ADP)ribose polymerase (PARP) are calpain substrates.
We demonstrate here that both procaspase-3 (32 kDa) and PARP are calpain substrates. In calcium-channel opener maitotoxin-treated cells, a 30 kDa caspase-3 fragment is produced in a time and concentration-dependent manner. Formation of this fragment is prevented by calpain inhibitors but not by the pancaspase inhibitor, carbobenzoxy-Asp-CH(2)OC(O)-2,6-dichlorobenzene (Z-D-DCB) nor the selective proteasome inhibitor lactacystin. In maitotoxin-treated cells, PARP (113 kDa) is also cleaved into a 40 kDa immunoreactive fragment, in a calpain-inhibitor-sensitive manner. Both procaspase-3 and PARP are also cleaved in vitro by purified micro-calpain to a 30 kDa fragment and a 40 kDa fragment, respectively. Finally, we show that staurosporine-mediated caspase-3 activation is interrupted by maitotoxin pretreatment.
McGinnis
K M
KM
Department of Pharmacology, University of Michigan Medical School, Ann Arbor, Michigan 48109, USA.
Gnegy
M E
ME
Park
Y H
YH
Mukerjee
N
N
Wang
K K
KK
D000111:Acetylcysteine / Q000031:analogs & derivatives / Q000494:pharmacology; D001224:Aspartic Acid / Q000031:analogs & derivatives / Q000494:pharmacology; D002154:Calpain / Q000378:metabolism*; D053148:Caspase 3; D061945:Caspase Inhibitors; D020169:Caspases / Q000737:chemistry / Q000378:metabolism*; D002460:Cell Line; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects; D004792:Enzyme Precursors / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D008387:Marine Toxins / Q000494:pharmacology; D035964:Oxocins; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism*; D011499:Protein Processing, Post-Translational / Q000187:drug effects; D019311:Staurosporine / Q000494:pharmacology; D013379:Substrate Specificity; D000095702:Polyether Toxins
D000111:Acetylcysteine; D001224:Aspartic Acid; D002154:Calpain; D053148:Caspase 3; D061945:Caspase Inhibitors; D020169:Caspases; D015853:Cysteine Proteinase Inhibitors; D004792:Enzyme Precursors; D008387:Marine Toxins; D035964:Oxocins; D011065:Poly(ADP-ribose) Polymerases; D019311:Staurosporine; C093260:benzyloxycarbonyl-Asp-CH2OC(O)-2,6-dichlorobenzene; C067713:lactacystin; C020394:maitotoxin; D000095702:Polyether Toxins; C505407:CASP3 protein, human