Kumar Sharad, Dorstyn Loretta
Hanson Institute, Adelaide, Australia.
Methods Mol Biol. 2009;559:3-17. doi: 10.1007/978-1-60327-017-5_1.
Apoptotic cell death is characterised by various morphological and biochemical changes. Cysteine proteases of the caspase family play key roles in the execution of apoptosis and in the maturation of proinflammatory cytokines. During apoptosis signalling, caspase precursors undergo rapid proteolytic processing and activation. Activated caspases then function to cleave various vital cellular proteins, resulting in the death of the cell. Thus, the measurement of caspase activation and caspase activity provides a quick and convenient method to assess apoptosis. This chapter outlines various commonly used assays for measuring caspase activity and detecting active caspases in cultured cells or tissue extracts.
凋亡性细胞死亡具有多种形态学和生物化学变化特征。半胱天冬酶家族的半胱氨酸蛋白酶在凋亡的执行以及促炎细胞因子的成熟过程中发挥关键作用。在凋亡信号传导过程中，半胱天冬酶前体经历快速的蛋白水解加工和激活。激活的半胱天冬酶随后发挥作用，切割各种重要的细胞蛋白，导致细胞死亡。因此，测量半胱天冬酶的激活和活性提供了一种快速便捷的方法来评估细胞凋亡。本章概述了用于测量培养细胞或组织提取物中半胱天冬酶活性和检测活性半胱天冬酶的各种常用检测方法。
