Zhivotovsky B, Samali A, Gahm A, Orrenius S
Division of Toxicology, Institute of Environmental Medicine, Karolinska Institutet, Box 210, S-171 77 Stockholm, Sweden.
Cell Death Differ. 1999 Jul;6(7):644-51. doi: 10.1038/sj.cdd.4400536.
The activation of the caspase family of proteases has been detected in numerous cell systems and appears to function as a common pathway through which apoptotic mechanisms may operate. Caspases are synthesized as precursors (pro-caspases) and are converted into mature enzymes by apoptotic signals. The effects of caspases in apoptosis are accomplished by the cleavage of numerous proteins located in different intracellular compartments. In the present study we have addressed the question of the subcellular localization of different pro- and active caspases as well as several other proteins, such as Apaf-1, calpain and DFF, which also play important roles in the apoptotic process. We found that at least three pro-caspases (pro-caspases-2, -3 and -9) were present in both the mitochondrial and cytosolic fractions of untreated Jurkat T lymphocytes. Only pro-caspase-2 was found in the nuclear fraction. Pro-caspases-7 and -8 were found only in the cytosolic fraction. In apoptotic cells, caspases-3, -8 and -9 were present in the cytosolic fraction, whereas caspases-3 and -9 were also found in the mitochondrial fraction and caspase-7 in the microsomal fraction. Caspases-2 and -3 were present in the nuclear fraction. The selective localization of pro-caspases in different subcellular compartments may play an important, but yet unknown, role in their activation. The translocation of active caspases to other subcellular compartments appears to be critical for the development of the apoptotic process.
在众多细胞系统中均检测到蛋白酶半胱天冬酶家族的激活,其似乎作为细胞凋亡机制可能发挥作用的共同途径。半胱天冬酶最初以前体(前半胱天冬酶)形式合成,并通过凋亡信号转化为成熟酶。半胱天冬酶在细胞凋亡中的作用是通过切割位于不同细胞内区室的众多蛋白质来实现的。在本研究中,我们探讨了不同前半胱天冬酶和活性半胱天冬酶以及其他几种蛋白质(如凋亡蛋白酶激活因子-1、钙蛋白酶和DNA片段化因子)的亚细胞定位问题,这些蛋白质在细胞凋亡过程中也发挥着重要作用。我们发现,至少三种前半胱天冬酶(前半胱天冬酶-2、-3和-9)存在于未处理的Jurkat T淋巴细胞的线粒体和胞质组分中。仅在前半胱天冬酶-2存在于核组分中。前半胱天冬酶-7和-8仅存在于胞质组分中。在凋亡细胞中,半胱天冬酶-3、-8和-9存在于胞质组分中,而半胱天冬酶-3和-9也存在于线粒体组分中,半胱天冬酶-7存在于微粒体组分中。半胱天冬酶-2和-3存在于核组分中。前半胱天冬酶在不同亚细胞区室中的选择性定位可能在其激活过程中发挥重要但尚不清楚的作用。活性半胱天冬酶向其他亚细胞区室的转位似乎对细胞凋亡过程的发展至关重要。