Affinity labeling displays the stepwise activation of ICE-related proteases by Fas, staurosporine, and CrmA-sensitive caspase-8
| Autor: | Richard W. Moyer, Hirokazu Hirata, Kokichi Yamamoto, Toshiro Okazaki, Hirofumi Sawai, Shuji Kishi, Peter W. Mesner, Kyung-Kwon Lee, Minoru Okuma, Shin Yonehara, Masataka Sasada, Atsushi Takahashi, Peter C. Turner, Yuzuru Imai |
|---|---|
| Rok vydání: | 1997 |
| Předmět: |
Cancer Research
Proteases Programmed cell death Apoptosis Cysteine Proteinase Inhibitors Caspase 8 Jurkat cells Substrate Specificity Jurkat Cells Viral Proteins Genetics medicine Animals Humans Staurosporine fas Receptor Molecular Biology Serpins Caspase Affinity labeling Caspase 6 biology Affinity Labels Molecular biology Caspase 9 Enzyme Activation Cysteine Endopeptidases Caspases biology.protein Laminin Poly(ADP-ribose) Polymerases Chickens Oligopeptides medicine.drug |
| Zdroj: | Oncogene. 14:2741-2752 |
| ISSN: | 1476-5594 0950-9232 |
| DOI: | 10.1038/sj.onc.1201131 |
| Popis: | The activation of multiple interleukin-1beta converting enzyme-related proteases (caspases) in apoptotic mammalian cells raises questions as to whether the multiple active caspases have distinct roles in apoptotic execution as well as how these proteases are organized in apoptotic signaling pathways. Here we used an affinity-labeling agent, YV(bio)KD-aomk, to investigate the caspases activated during apoptotic cell death. YV(bio)KD-aomk identified six distinct polypeptides corresponding to active caspases in Fas-stimulated Jurkat T cells. On staurosporine treatment, four polypeptides were detected. Competition experiments showed that the labeled caspases have distinct substrate preferences. Stepwise appearance of the labeled caspases in each cell death event was consistent with the view that the activated caspases are organized into protease cascades. Moreover, we found that stepwise activation of caspases similar to that induced by Fas ligation is triggered by exposing non-apoptotic Jurkat cell extracts to caspase-8 (MACH/FLICE/Mch5). Conversely, CrmA protein, a viral suppressor of Fas-induced apoptosis, inhibited the protease activity of caspase-8. Overall, these findings provide evidence that caspase-8, a CrmA-sensitive protease, is responsible for initiating the stepwise activation of multiple caspases in Fas-stimulated cells. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|