Köhler Camilla, Orrenius Sten, Zhivotovsky Boris
Division of Toxicology, Institute of Environmental Medicine, Karolinska Institutet, Box 210, SE-171 77 Stockholm, Sweden.
J Immunol Methods. 2002 Jul 1;265(1-2):97-110. doi: 10.1016/s0022-1759(02)00073-x.
A family of cysteine proteases, the caspases, plays a central role in the initiation and execution phases of apoptosis. Upon activation, these enzymes cleave specific substrates and thereby mediate many of the typical biochemical and morphological changes in apoptotic cells, such as cell shrinkage, chromatin condensation, DNA fragmentation and plasma membrane blebbing. Hence, the detection of activated caspases can be used as a biochemical marker for apoptosis. Here we review a set of methods available for characterizing and quantifying the activation of caspases, including immunoblotting, cleavage of synthetic substrates, affinity labeling and confocal microscopy. Each method is described in general terms and the advantages and disadvantages of each technique are discussed.
半胱天冬酶家族，即胱天蛋白酶，在细胞凋亡的起始和执行阶段起着核心作用。激活后，这些酶会切割特定底物，从而介导凋亡细胞中许多典型的生化和形态学变化，如细胞皱缩、染色质凝聚、DNA片段化和质膜起泡。因此，检测活化的胱天蛋白酶可作为细胞凋亡的生化标志物。在此，我们综述了一系列可用于表征和定量胱天蛋白酶激活的方法，包括免疫印迹法、合成底物切割法、亲和标记法和共聚焦显微镜法。每种方法均以一般术语进行描述，并讨论了每种技术的优缺点。
