若丹明110连接的氨基酸和肽作为底物，用于测量完整细胞凋亡诱导时的半胱天冬酶活性。

Rhodamine 110-linked amino acids and peptides as substrates to measure caspase activity upon apoptosis induction in intact cells.

作者信息

Hug H, Los M, Hirt W, Debatin K M

机构信息

Universitäts-Kinderklinik Ulm, Germany.

出版信息

Biochemistry. 1999 Oct 19;38(42):13906-11. doi: 10.1021/bi9913395.

DOI:10.1021/bi9913395

PMID:10529236

Abstract

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.

摘要

半胱天冬酶（天冬氨酸特异性半胱氨酸蛋白酶）是一组结构相关的半胱氨酸蛋白酶，它们在特定识别序列后切割肽键。它们在激活脊椎动物细胞凋亡中起核心作用。为了测量各种刺激诱导的凋亡以及在凋亡早期阶段，半胱天冬酶是一个理想的靶点。当必须在吞噬细胞清除凋亡细胞之前对其进行体外分析时，情况尤其如此。一种新的灵敏的半胱天冬酶检测方法基于一种仅含有与罗丹明110相连的天冬氨酸残基的底物。使用这种底物和类似的底物，我们能够在完整造血细胞系中诱导凋亡后通过流式细胞术检测细胞内半胱天冬酶的激活。