| Autor: |
Margaret M, Racke, Marian, Mosior, Steve, Kovacevic, Chan Hsin S, Chang, Andrew L, Glasebrook, Neal W, Roehm, Songqing, Na |
| Rok vydání: |
2002 |
| Předmět: |
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| Zdroj: |
Journal of neurochemistry. 80(6) |
| ISSN: |
0022-3042 |
| Popis: |
Activated caspase-3 is considered an important enzyme in the cell death pathway. To study the specific role of caspase-3 activation in neuronal cells, we generated a stable tetracycline-regulated SK-N-MC neuroblastoma cell line, which expressed a highly efficient self-activating chimeric caspase-3, consisting of the caspase-1 prodomain fused to the caspase-3 catalytic domain. Under expression-inducing conditions, we observed a time-dependent increase of processed caspase-3 by immunostaining for the active form of the enzyme, intracellular caspase-3 enzyme activity, as well as poly(ADP-ribose) polymerase (PARP) cleavage. Induced expression of the caspase fusion protein showed predominantly caspase-3 activity without any apoptotic morphological changes. In contrast, staurosporine treatment of the same cells resulted in activation of multiple caspases and profound apoptotic morphology. Our work provides evidence that auto-activation of caspase-3 can be efficiently achieved with a longer prodomain and that neuronal cell apoptosis may require another caspase or activation of multiple caspase enzymes. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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