Identification of a caspase-9 substrate and detection of its cleavage in programmed cell death during mouse development.
Identification of a caspase-9 substrate and detection of its cleavage in programmed cell death during mouse development.
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DOI：
10.1074/jbc.m105648200
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发表时间：
2001-11-02
影响因子：
4.8
通讯作者：
Morishima, N
中科院分区：
文献类型：
--
作者：
Nakanishi, K;Maruyama, M;Morishima, N
caspase家族的蛋白酶代表了细胞凋亡发生的主要机制。体外研究表明，上游caspase在响应凋亡刺激时被激活，而活性caspase反过来又加工下游效应caspase，参与细胞结构的破坏。caspase -9是一种上游caspase，可在细胞损伤反应中变得活跃，包括体外生长因子剥夺和暴露于氧化应激。然而，关于caspase-9的激活如何在体内（例如在发育过程中）受到时间和空间调节，我们知之甚少。我们已经确定了vimentin作为caspase-9底物的第一个例子，它不是下游的procaspase。免疫组化分析显示，caspase-9可在胚胎神经系统和指间区凋亡细胞中切割vimentin。这一结果与caspase-9及其激活因子Apaf-1基因敲除导致这些组织发育缺陷的观察结果一致。我们的研究结果表明，特异性抗体可用于原位检测程序性细胞死亡中caspase-9的激活。
The caspase family of proteases represents the main machinery by which apoptosis occurs. In vitro studies have revealed that upstream caspases are activated in response to apoptotic stimuli, and the active caspases in turn process downstream effector caspases that are involved in the destruction of cellular structure. Caspase-9 is an upstream caspase that can become active in response to cellular damage, including deprivation of growth factors and exposure to oxidative stress in vitro. Little is known, however, about how activation of caspase-9 is temporally and spatially regulated in vivo, e.g. during development. We have identified vimentin as the first example of a caspase-9 substrate that is not a downstream procaspase. Immunohistochemical analysis, using a specific antibody against the vimentin fragments generated by caspase-9, showed that caspase-9 cleaves vimentin in apoptotic cells in the embryonic nervous system and the interdigital regions. This result is consistent with observations that gene knockouts of caspase-9 and its activator, Apaf-1, result in developmental defects in these tissues. Our results show that the specific antibody is useful for in situ detection of caspase-9 activation in programmed cell death.