Involvement of cytoplasmic serine proteinase and CPP32 subfamily in the molecular machinery of caspase 3 activation during Fas-mediated apoptosis.
On stimulation by the Fas ligand, the death receptor, Fas, initiates a signal leading to apoptotic cell death. Fas plays an important role in physiological cell death and is closely involved in various disease states. Recent investigations have shown that caspase 3 plays a dominant role in the process of Fas-mediated apoptosis. In the present study, we investigated the molecular machinery of caspase 3 activation in Fas-mediated apoptosis. The results showed that Fas-mediated apoptosis was accompanied by caspase 3 activation, and both Fas-mediated apoptosis and caspase 3 activation were prevented by a serine proteinase inhibitor. In addition, the serine proteinase inhibitor also prevented the caspase 3 activation in cytoplasts, and the specific activation of serineproteinase was encountered in only cytoplasmic proteins. These results suggest that cytoplasmic serineproteinase plays an important role in caspase 3 activation. Interestingly, caspase 3 was cleaved at p3 site immediately after Fas Ab stimulation, and the cleavage at p17 site became detectable later. We also found that among tested proteinases only Staphylococcus aureus V8 serineproteinase initiated caspase 3 activation and specifically cleaved at p3 site. These results strongly suggest that a cytoplasmic S. aureus V8-like serine proteinase is closely involved in caspase 3 activation.
Suzuki
A
A
Drug Safety Research Laboratory, Daiichi Pharmaceutical Co., Ltd, Tokyo R&D center, Edogawa-ku, Japan. LEB00373@niftyserve.or.jp
Iwasaki
M
M
Wagai
N
N
D017209:Apoptosis; D053148:Caspase 3; D020169:Caspases; D002451:Cell Compartmentation; D003546:Cysteine Endopeptidases / Q000378:metabolism*; D003593:Cytoplasm / Q000201:enzymology; D004789:Enzyme Activation; D053222:Fas Ligand Protein; D006367:HeLa Cells; D006801:Humans; D008562:Membrane Glycoproteins / Q000378:metabolism*; D009842:Oligopeptides / Q000494:pharmacology; D011480:Protease Inhibitors / Q000494:pharmacology; D012697:Serine Endopeptidases / Q000378:metabolism; D015398:Signal Transduction; D019014:fas Receptor / Q000378:metabolism*
C505661:FASLG protein, human; D053222:Fas Ligand Protein; D008562:Membrane Glycoproteins; D009842:Oligopeptides; D011480:Protease Inhibitors; C099158:acetyl-aspartyl-glutamyl-valyl-aspartal; D019014:fas Receptor; D012697:Serine Endopeptidases; C505407:CASP3 protein, human; D053148:Caspase 3; D020169:Caspases; D003546:Cysteine Endopeptidases