The three-dimensional structure of apopain/CPP32, a key mediator of apoptosis

Rotonda, J.; Nicholson, Donald W.; Fazil, Kimberly M.; Gallant, Michel; Gareau, Yves; Labelle, Marc; Peterson, Erin P.; Rasper, Dita; Ruel, Réjean; Vaillancourt, John P.; Thornberry, Nancy A.; Becker, Joseph W.

doi:10.1038/nsb0796-619

Cited by 415 publications

(381 citation statements)

References 36 publications

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“…The mature and active form of all caspases is a heterotetrameric complex composed of two large subunits and two small subunits. 75,76 Caspases with large prodomains, eg caspase-2, -8 and -9, are thought to be involved in the initiation of the apoptotic response and are therefore called initiator caspases. Initiator caspases are directly linked to different death-inducing signaling complexes, such as the CD95/Fas signaling complex (pro-caspase-8) or the mitochondrial apoptosome (pro-caspase-9), by protein interaction motifs in their prodomains.…”

Section: Initiator and Effector Caspases And Their Targetsmentioning

confidence: 99%

The kiss of death: promises and failures of death receptors and ligands in cancer therapy

et al. 2001

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Section: Initiator and Effector Caspases And Their Targetsmentioning

confidence: 99%

The kiss of death: promises and failures of death receptors and ligands in cancer therapy

et al. 2001

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“…The X-ray crystal structure of active caspase-3 has been solved. 14,15 The mature enzyme was shown to exist as a tetramer with two independent active sites, each containing elements from one large (p17) and one small (p12) subunit. Catalysis is mediated by a mechanism typical of cysteine proteases involving a catalytic dyad, composed of Cys163 and His121, which are harbored by the large subunit.…”

Section: Introductionmentioning

confidence: 99%

“…The substrate-binding cleft recognizes a short 4 amino-acid stretch within protein substrates, directly N-terminal to the cleavage site. 14 This tetrapeptide motif, which is sufficient to bind specifically to the active caspase, is the basis for the design of the synthetic inhibitors. Comparative inhibitor analysis clearly indicates that peptide-derived inhibitors harboring the optimal substrate tetrapeptide-motif for a specific caspase are highly potent and in some cases selective.…”

Section: Introductionmentioning

confidence: 99%

Catalytic activity of caspase-3 is required for its degradation: stabilization of the active complex by synthetic inhibitors

et al. 2004

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The activation of caspase-3 represents a critical step in the pathways leading to the biochemical and morphological changes that underlie apoptosis. Upon induction of apoptosis, the large (p17) and small (p12) subunits, comprising active caspase-3, are generated via proteolytic processing of a latent proenzyme dimer. Two copies of each individual subunit are generated to form an active heterotetramer. The tetrameric form of caspase-3 cleaves specific protein substrates within the cell, thereby producing the apoptotic phenotype. In contrast to the proenzyme, once activated in HeLa cells, caspase-3 is difficult to detect due to its rapid degradation. Interestingly, however, enzyme stability and therefore detection of active caspase-3 by immunoblot analysis can be restored by treatment of cells with a peptide-based caspase-3 selective inhibitor, suggesting that the active form can be stabilized through protein-inhibitor interaction. The heteromeric active enzyme complex is necessary for its stabilization by inhibitors, as expression of the large subunit alone is not stabilized by the presence of inhibitors. Our results show for the first time, that synthetic caspase inhibitors not only block caspase activity, but may also increase the stability of otherwise rapidly degraded mature caspase complexes. Consistent with these findings, experiments with a catalytically inactive mutant of caspase-3 show that rapid turnover is dependent on the activity of the mature enzyme. Furthermore, turnover of otherwise stable active site mutants of capase-3 is rescued by the presence of the active enzyme suggesting that turnover can be mediated in trans.

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“…Crystal structures of caspase-1 (ICE) (Walker et al, 1994;Wilson et al, 1994) and caspase-3 (CPP32/Yama/apopain) (Rotonda et al, 1996) suggest that active forms of caspases are tetramers composed of two pairs of large and small subunits, which are derived from two molecules of a precursor by cleavage at multiple Asp-X bonds. This unusual proteolytic processing suggests that caspase activation is driven by autocatalysis, or by cleavage by other caspases (Martin and Green, 1995).…”

Section: Introductionmentioning

confidence: 99%

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

et al. 1997

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The activation of multiple interleukin-1b 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 a nity-labeling agent, YV(bio)KD-aomk, to investigate the caspases activated during apoptotic cell death. YV(bio)KD-aomk identi®ed 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 ®ndings provide evidence that caspase-8, a CrmA-sensitive protease, is responsible for initiating the stepwise activation of multiple caspases in Fas-stimulated cells.

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The three-dimensional structure of apopain/CPP32, a key mediator of apoptosis

Cited by 415 publications

References 36 publications

The kiss of death: promises and failures of death receptors and ligands in cancer therapy

The kiss of death: promises and failures of death receptors and ligands in cancer therapy

Catalytic activity of caspase-3 is required for its degradation: stabilization of the active complex by synthetic inhibitors

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

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