Rhodamine 110-linked amino acids and peptides as substrates to measure caspase activity upon apoptosis induction in intact cells
Rhodamine 110-linked amino acids and peptides as substrates to measure caspase activity upon apoptosis induction in intact cells
复制标题
DOI：
10.1021/bi9913395
复制
发表时间：
1999-10-19
期刊：
影响因子：
2.9
通讯作者：
Debatin, KM
中科院分区：
文献类型：
--
作者：
Hug, H;Los, M;Debatin, KM
半胱氨酸蛋白酶（半胱氨酸天冬氨酸特异性蛋白酶）是一组结构相关的半胱氨酸蛋白酶，其切割特定识别序列后的肽键。它们在激活脊椎动物细胞的凋亡中起核心作用。为了测量由各种刺激诱导的凋亡和在早期凋亡阶段，半胱天冬酶是理想的靶标。当凋亡细胞必须在吞噬细胞去除它们之前进行离体分析时，情况尤其如此。一种新的和敏感的半胱天冬酶测定是基于一个底物，只含有天冬氨酸残基连接到罗丹明110。与此类似的基板，我们能够检测细胞内的半胱天冬酶激活后，在完整的造血细胞系凋亡诱导的流式细胞术。
Caspases (cysteine aspartate-specific proteases) are a structurally related group of cysteine proteases that cleave peptide bonds following specific recognition sequences. They play a central role in activating apoptosis of vertebrate cells. To measure apoptosis induced by various stimuli and at an early apoptotic stage, caspases are an ideal target. This is especially the case when apoptotic cells have to be analyzed ex vivo before phagocytes remove them. A new and sensitive caspase assay is based on a substrate that contains only aspartate residues linked to rhodamine 110. With this and similar substrates, we are able to detect intracellular caspase activation by flow cytometry after apoptosis induction in intact hematopoetic cell lines.