Affinity labeling displays the stepwise activation of ICE-related proteases by Fas, staurosporine, and CrmA-sensitive caspase-8

被引:102
作者
Takahashi, A
Hirata, H
Yonehara, S
Imai, Y
Lee, KK
Moyer, RW
Turner, PC
Mesner, PW
Okazaki, T
Sawai, H
Kishi, S
Yamamoto, K
Okuma, M
Sasada, M
机构
[1] KYOTO UNIV,DEPT HEMATOL & ONCOL,GRAD SCH MED,KYOTO 606,JAPAN
[2] KYOTO UNIV,DEPT VIRAL ONCOL,INST VIRUS RES,KYOTO 606,JAPAN
[3] UNIV FLORIDA,COLL MED,DEPT MOL GENET & MICROBIOL,GAINESVILLE,FL 32610
[4] MAYO CLIN & MAYO FDN,DIV ONCOL RES,ROCHESTER,MN 55905
[5] JAPAN TOBACCO INC,PHARMACEUT BASIC RES LABS,YOKOHAMA,KANAGAWA 236,JAPAN
[6] KYOTO UNIV,COLL MED TECHNOL,KYOTO 606,JAPAN
关键词
apoptosis; apoptotic cell death; YV(bio)KD-aomk; serpin; Jurkat T cells;
D O I
10.1038/sj.onc.1201131
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The activation of multiple interleukin-1 beta 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 sis distinct polypeptides corresponding to active caspases in Fas-stimulated Jurkat T cells, On staurosporine treatment, four polypeptides were detected, Competition experiments showed that the labelled caspases have distinct substrate preferences, Stepwise appearance of the labelled caspases in each cell death event was consistent with the view that the activated caspases are organized into protease cascades, Moreover, me 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.
引用
收藏
页码:2741 / 2752
页数:12
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